Spirocyclic compounds as farnesoid x receptor modulators

ABSTRACT

The present invention provides compounds of Formula (I): 
     
       
         
         
             
             
         
       
     
     or stereoisomers, tautomers, or pharmaceutically acceptable salts or solvates thereof, wherein all the variables are as defined herein. These compounds modulate the activity of famesoid X receptor (FXR), for example, as agonists. This invention also relates to pharmaceutical compositions comprising these compounds and methods of treating a disease, disorder, or condition associated with FXR dysregulation, such as pathological fibrosis, transplant rejection, cancer, osteoporosis, and inflammatory disorders, by using the compounds and pharmaceutical compositions.

CROSS REFERENCE

This application claims the benefit of U.S. Provisional Application Ser.No. 62/580,068 filed Nov. 1, 2017 which is incorporated herein in itsentirety.

DESCRIPTION

The present invention relates generally to compounds useful as famesoidX receptor (FXR) modulators, pharmaceutical compositions comprising suchcompounds and to their use in therapy, especially in the treatment orprophylaxis of diseases, disorders, and conditions for which an FXRmodulator is indicated.

FXR or NR1H4 (nuclear receptor subfamily 1, group H, member 4) is anuclear receptor that can activate the expression of specific targetgenes in a ligand-dependent manner. FXR is expressed in the liver,throughout the gastrointestinal tract, colon, ovary, adrenal gland,kidney, and in the gall bladder and biliary tree in humans. FXR forms aheterodimer with Retinoid X Receptor (RXR) and binds to specificresponse elements in target genes to regulate gene transcription (B. M.Forman et al., Cell 1995; 81: 687; W. Seol et al., Mol. Endocrinol.1995; 9: 72). The FXR/RXR heterodimer typically binds to an invertedrepeat of a consensus hexanucleotide sequence (AGGTCA) separated by asingle nucleotide, i.e. an IR-1 sequence. The relevant physiologicalligands of FXR are bile acids including chenodeoxycholic acid and itstaurine-conjugate (D. J. Parks et al., Science 1999; 284: 1365; M.Makishima et al., Science 1999; 284: 1362). FXR activation regulates theexpression of multiple genes that encode enzymes and transportersinvolved in bile acid synthesis, influx, and efflux from the liver andintestine resulting in a net decrease in total endogenous bile acids ina negative feedback loop. FXR is involved in paracrine and endocrinesignaling by upregulating the expression of the cytokine FibroblastGrowth Factor 15 (rodents) or 19 (primates), which can also contributeto the regulation of bile acid concentrations (Holt et al., Genes Dev.2003; 17: 1581; Inagaki et al., Cell Metab 2005; 2: 217). Therefore, FXRis considered to be a master regulator of bile acid homeostasis.

One use of FXR agonists is for the treatment of diseases in which bileacids are dysregulated, including cholestatic diseases (e.g. primarybiliary cirrhosis and primary sclerosing cholangitis) that can lead tofibrosis, cirrhosis, cholangiocarcinoma, hepatocellular carcinoma, liverfailure, and death. While elevated bile acid concentrations in the liverhave deleterious effects, bile acids also affect the microflora andintegrity of the small intestine. Obstruction of bile flow in humans orrodents causes proliferation of intestinal bacteria and mucosal injury,which can lead to bacterial translocation across the mucosal barrier andsystemic infection (Berg, Trends Microbiol. 1995; 3: 149-154). Micelacking FXR have increased ileal levels of bacteria and a compromisedepithelial barrier, while activation of intestinal FXR plays animportant role in preventing bacterial overgrowth and maintaining theintegrity of the intestinal epithelium (Inagaki et al., Proc Natl AcadSci 2006; 103: 3920-3925). Over time, FXR null mice spontaneouslydevelop hepatocellular carcinoma, and this can be abrogated by selectivere-activation of FXR in the intestine (Degirolamo et al., Hepatology 61:161-170). Pharmacological activation of FXR with a small moleculeagonist or transgenic expression of FXR in the intestine can normalizebile acid concentrations, decrease cellular proliferation in hepaticbile ducts, and reduce inflammatory cell infiltration, necrotic area,and liver fibrosis in rodent models of cholestasis (Liu et al., J. Clin.Invest. 2003; 112:1678-1687; Modica et al., Gastroenterology. 2012; 142:355-365). Some of these beneficial effects observed in preclinicalmodels of cholestasis have translated to human patients, and the FXRagonist, obeticholic acid (OCA or OCALIVA™), has been approved for thetreatment of primary biliary cirrhosis(https://www.fda.gov/newsevents/newsroom/pressannouncements/ucm503964.htm).

In addition to controlling bile acid homeostasis, FXR agonists regulatethe hepatic expression of hundreds of genes encoding proteins involvedin cholesterol and lipid metabolism and transport, glucose homeostasis,inflammation, chemotaxis, and apoptosis among other pathways (Zhan etal., PLoS One 2014; 9: e105930; Ijssennagger et al., J Hepatol 2016; 64:1158-1166). Consistent with these broad effects on gene expression, FXRagonists have also been investigated in preclinical models of fibrosis,cancer, inflammatory diseases, and metabolic disorders, includingdyslipidemia, obesity, type 2 diabetes, nonalcoholic fatty liver disease(NAFLD) and metabolic syndrome (Crawley, Expert Opin. Ther. Patents2010; 20:1047-1057).

FXR agonists are also being investigated in human clinical trials forthe treatment of NAFLD, a more advanced form of fatty liver disease,nonalcoholic steatohepatitis (NASH), and associated complications. NAFLDis one of the most common causes of chronic liver disease in the worldtoday (Vemon et al., Aliment Pharmacol Ther 2011; 34:274-285). The riskfactors for developing NAFLD include obesity, type 2 diabetes mellitus(T2DM), insulin resistance, hypertension, and dyslipidemia. In a 6-weekclinical trial in T2DM patients with NAFLD, the FXR agonist OCAstatistically significantly improved insulin sensitivity and reducedbody weight, showing beneficial effects on some of these risk factors(Mudaliar et al., Gastroenterology 2013; 145: 574-582). NASH is the mostsevere and progressive form of NAFLD and includes the histologicalfindings of hepatic steatosis, inflammation, and ballooning degenerationwith varying amounts of pericellular fibrosis (Sanyal et al., Hepatology2015; 61:1392-1405). In a 72-week clinical trial in patients with NASH,OCA statistically significantly improved hepatic steatosis, lobularinflammation, hepatocyte ballooning, and fibrosis as assessed byhistological analyses of liver biopsies (Neuschwander-Tetri et al.,Lancet 2015; 385: 956-965). These data also suggest the potential forFXR agonists to show benefit on clinical outcomes given that NASH is thesecond leading cause of hepatocellular carcinoma (HCC) and livertransplantation in the United States (Wong et al., Hepatology 2014; 59:2188-2195).

The present invention provides novel compounds for treating a disease,disorder, or condition associated with famesoid X receptor (FXR)activity in a patient in need thereof.

SUMMARY OF THE INVENTION

In one aspect, the present invention provides compounds of Formula (I),Formula (II) and Formula (III) as well as the subgenera and speciesthereof, including stereoisomers, tautomers, pharmaceutically acceptablesalts, and solvates thereof, which are useful as FXR modulators.

In another aspect, the present invention also provides processes andintermediates for making the compounds of the present invention.

In another aspect, the present invention also provides pharmaceuticalcompositions comprising a pharmaceutically acceptable carrier and atleast one of the compounds of the present invention or stereoisomers,tautomers, pharmaceutically acceptable salts, or solvates thereof.

In another aspect, the compounds of the invention may be used intherapy, either alone or in combination with one or more additionaltherapeutic agents.

The compounds of the invention may be used in the treatment of adisease, disorder, or condition associated with activity of famesoid Xreceptor (FXR) in a patient in need of such treatment by administering atherapeutically effective amount of the compound, or a stereoisomer, atautomer, or a pharmaceutically acceptable salt or solvate thereof, tothe patient. The disease, disorder, or condition may be related topathological fibrosis. The compounds of the invention can be used alone,in combination with one or more compounds of the present invention, orin combination with one or more, e.g., one to two, other therapeuticagents.

The compounds of the invention may be used, either as a single agent orin combination with other agents, in the treatment of a disease,disorder, or condition selected from nonalcoholic steatohepatitis(NASH), non-alcoholic fatty liver disease (NAFLD), chronic kidneydisease, diabetic kidney disease, primary sclerosing cholangitis (PSC),and primary biliary cirrhosis (PBC). The compounds of the invention maybe used, either as a single agent or in combination with other agents,in the treatment of idiopathic pulmonary fibrosis (IPF).

The compounds of the invention may be used for the manufacture of amedicament for the treatment of a disease, disorder, or condition in apatient in need of such treatment.

Other features and advantages of the invention will be apparent from thefollowing detailed description and claims.

DETAILED DESCRIPTION

The present application provides compounds, including all stereoisomers,solvates, prodrugs and pharmaceutically acceptable salt and solvateforms thereof, according to Formula (I). The present application alsoprovides pharmaceutical compositions containing at least one compoundaccording to Formula (I), or a stereoisomer, a tautomer, or apharmaceutically acceptable salt or a solvate thereof, and optionally atleast one additional therapeutic agent. Additionally, the presentapplication provides methods for treating a patient suffering from aFXR-modulated disease or disorder such as for example, biliary fibrosis,liver fibrosis, renal fibrosis, Non-Alcoholic Fatty Liver Disease(NAFLD), Non-Alcoholic Steato-Hepatitis (NASH), primary sclerosingcholangitis (PSC), primary biliary cirrhosis (PBC), and pancreaticfibrosis, by administering to a patient in need of such treatment atherapeutically effective amount of a compound of the present invention,or a stereoisomer, a tautomer, or a pharmaceutically acceptable salt ora solvate thereof, and optionally in combination with at least oneadditional therapeutic agent.

I. Compounds of the Invention

In one embodiment, the present invention provides a compound of Formula(I):

-   X¹ and X⁴ are each independently C or N;-   X² and X³ are each independently CR⁵, N, NR⁶, O, or S;-   E ring is a 4- to 6-membered carbocyclyl or heterocyclyl, wherein    the carbocyclyl and heterocyclyl are each independently substituted    with 0 to 3 R³;-   * denotes a spiro carbon atom;-   Y is CR⁷ or N;-   m and n are each independently an integer of 0, 1, or 2;-   f is an integer of 0, 1, 2, or 3;-   Z is 6- to 10-membered aryl, 5- to 10-membered heteroaryl containing    1 to 3 heteroatoms independently selected from N, O, and S, 3- to    10-membered carbocyclyl, or 4- to 10-membered heterocyclyl    containing 1 to 3 heteroatoms independently selected from N, O, and    S, wherein the aryl, heteroaryl carbocyclyl, and heterocyclyl are    independently substituted with 0 to 5 R⁸;-   L¹ is a covalent bond, O, S, NR¹⁶, —S(O)₂—, C₁₋₃ alkylene, C₁₋₃    heteroalkylene, C₂₋₄ alkenylene, C₂₋₄ alkynylene, aryl, or a 5- to    6-membered heteroaryl containing 1 to 3 heteroatoms independently    selected from N, O, and S containing 1 to 4 heteroatoms    independently selected from N, O, and S; wherein the alkylene,    alkenylene, aryl, heteroalkylene, and heteroaryl are each    independently substituted with 0 to 3 R¹¹;-   L² is a covalent bond, O, S, NR¹⁷, C₁₋₃ alkylene, or C₁₋₃    heteroalkylene, wherein the alkylene and heteroalkylene are    independently substituted with 0 to 3 R¹⁵;-   R^(X) is -L³-R;-   L³ is a covalent bond, a C₁₋₃ alkylene, —C(O)NR¹²—CH₂—, or —OCH₂—,    wherein the C₁₋₃ alkylene is substituted with 0 to 3 R⁴;-   R^(Z) is —CN, —C(O)OR¹³, —C(O)NR^(14a)R^(14b),

-   R^(e) is C₁₋₆ alkyl, C₃₋₆ cycloalkyl, haloalkyl, hydroxyalkyl,    aminoalkyl, alkoxyalkyl, or haloalkoxyalkyl;-   each R^(Y) is independently hydrogen, halo, cyano, hydroxyl, amino,    C₁₋₆ alkyl, alkylamino, haloalkyl, hydroxyalkyl, aminoalkyl,    alkoxyalkyl, haloalkoxyalkyl, alkoxy, or haloalkoxy; or    alternatively two R^(Y), together with the carbon atoms to which    they are attached, form a bridge moiety; and with the proviso that    when Y is N and R^(Y) is attached to a carbon atom adjacent to Y,    then R^(Y) is not halo, cyano, hydroxyl, amino, alkoxy, or    haloalkoxy;-   R¹ is C₁₋₆ alkyl, C₃₋₅ cycloalkyl, or C₄₋₆ heterocyclyl, wherein the    alkyl or cycloalkyl is substituted with 0 to 3 R⁹;-   R² is 6- to 10-membered aryl, 5- to 10-membered heteroaryl    containing 1 to 3 heteroatoms independently selected from N, O, and    S, 3- to 10-membered carbocyclyl, or 4- to 10-membered heterocyclyl    containing 1 to 3 heteroatoms independently selected from N, O, and    S, wherein the aryl, heteroaryl, carbocyclyl, and heterocyclyl are    independently substituted with 0 to 5 R¹⁰;-   R³, R⁵ and R⁷ are each independently hydrogen, halo, cyano,    hydroxyl, amino, C₁₋₆ alkyl, alkylamino, haloalkyl, hydroxyalkyl,    aminoalkyl, alkoxyalkyl, haloalkoxyalkyl, alkoxy, or haloalkoxy;-   R⁴ is each independently halo, oxo, cyano, hydroxyl, amino, alkyl,    alkoxy, or alkylamino; or alternatively, two R⁴, taken together with    the atom(s) to which they are attached, form a carbocyclyl or    heterocyclyl moiety;-   R⁶, R¹⁶ and R¹⁷ are each independently hydrogen, C₁₋₆ alkyl,    haloalkyl, hydroxyalkyl, aminoalkyl, alkoxyalkyl, or    haloalkoxyalkyl;-   R⁸ and R¹⁰ are each independently halo, cyano, hydroxyl, amino, oxo,    —OR^(a), —SR^(a), ═S, —NR^(c)R^(c), ═NH, ═N—OH, ═NR^(a), ═N—OR^(a),    —NO₂, —S(O)₂R^(a), —S(O)₂NHR^(b), —S(O)₂NR^(c)R^(c), —S(O)₂OR^(b),    —OS(O)₂R^(b), —OS(O)₂OR^(b), —P(O)(OR^(b))(OR^(b)), —C(O)R^(b),    —C(NR^(b))R^(b), —C(O)OR^(b), —C(O)NR^(c)R^(c),    —C(NR^(b))NR^(c)R^(c), —OC(O)R^(b), —NR^(b)C(O)R^(b), —OC(O)OR^(b),    —NR^(b)C(O)OR^(b), —OC(O)NR^(c)R^(c), —NR^(b)C(O)NR^(c)R^(c),    —NR^(b)C(NR^(b))R^(b), —NR^(b)C(NR^(b))NR^(c)R^(c), C₁₋₆ alkyl, C₁₋₆    haloalkyl, aryl, arylalkyl, heteroaryl, carbocyclyl, or    heterocyclyl; wherein the alkyl, aryl, heteroaryl, carbocyclyl, and    heterocyclyl, by themselves or as part of another group, are each    independently substituted with 0 to 5 R^(d);-   R^(a) is each independently C₁₋₆ alkyl, haloalkyl, hydroxyalkyl,    aminoalkyl, alkoxyalkyl, haloalkoxyalkyl, aryl, arylalkyl,    heteroaryl, heteroarylalkyl, carbocyclyl, carbocyclylalkyl,    heterocyclyl, or heterocyclylalkyl;-   R^(b) is each independently hydrogen or R^(a);-   R^(c) is each independently R^(b) or alternatively, the two R^(c)    are taken together with the nitrogen atom to which they are bonded    form a 4-, 5-, 6- or 7-membered heterocyclyl containing 1 to 3    heteroatoms independently selected from N, O, and S;-   R^(d) is each independently selected from R^(a), alkoxy, haloalkoxy,    alkylamino, cycloalkylamino, heterocyclylamino, haloalkyl,    hydroxyalkyl, aminoalkyl, cycloalkoxy, heterocyclyloxy, haloalkoxy,    alkoxyalkoxy, haloalkylamino, alkoxyalkylamino,    haloalkoxyalkylamino, arylamino, aralkylamino, aryloxy, aralkyloxy,    heteroaryloxy, heteroarylalkyloxy, alkylthio, halo, cyano, hydroxyl,    amino, oxo, —OR^(a), —SR^(a), ═S, —NR^(c)R^(c), ═NH, ═N—OH, ═NR^(a),    ═N—OR^(a), —NO₂, —S(O)₂R^(a), —S(O)₂NHR^(b), —S(O)₂NR^(c)R^(c),    —S(O)₂OR^(b), —OS(O)₂R^(b), —OS(O)₂OR^(b), —P(O)(OR^(b))(OR^(b)),    —C(O)R^(b), —C(NR^(b))R^(b), —C(O)OR^(b), —C(O)NR^(c)R^(c),    —C(NR^(b))NR^(c)R^(c), —OC(O)R^(b), —NR^(b)C(O)R^(b), —OC(O)OR^(b),    —NR^(b)C(O)OR^(b), —NR^(b)C(O)NR^(c)R^(c), —NR^(b)C(NR^(b))R^(b),    and —NR^(b)C(NR^(b))NR^(c)R^(c);-   R⁹ is each independently halo, cyano, hydroxyl, amino, or C₁₋₆    alkyl;-   R¹¹ and R¹⁵ are each independently halo, oxo, cyano, hydroxyl,    amino, C₁₋₆ alkyl, C₃₋₆ cycloalkyl, C₄₋₆ heterocyclyl, alkylamino,    haloalkyl, hydroxyalkyl, aminoalkyl, alkoxyalkyl, haloalkoxyalkyl,    alkoxy, or haloalkoxy;-   R¹² is hydrogen or C₁₋₄ alkyl;-   R¹³ is hydrogen, C₁₋₁₀ alkyl, glycosyl, or    carboxy(trihydroxy)tetrahydropyranyl; and-   R^(14a) and R^(14b) are each independently hydrogen, C₁₋₆ alkyl,    C₃₋₆ cycloalkyl, C₄₋₆ heterocyclyl, alkylamino, haloalkyl,    hydroxyalkyl, aminoalkyl, alkoxyalkyl, haloalkoxyalkyl, alkoxy, or    haloalkoxy.

It should be understood by one skilled in the art that the dashed circledenotes an aromatic ring formed by X¹, X², X³, X⁴, and the carbon atom;and the dashed straight lines are each independently an optionalcovalent bond.

In one embodiment of Formula (I), X² is N or NR⁶.

In one embodiment of Formula (I), two R^(Y), together form a C₁₋₃alkylene bridge moiety. (R^(Y))_(f) denotes one or more optionalsubstituent groups on any of the suitable ring member atoms, and each ofR^(Y) is independent and can be the same or different.

In any one of the preceding embodiments of Formula (I), the

moiety is

In any one of the preceding embodiments of Formula (I), L¹ is a covalentbond, O, S, NH, C₁₋₃ alkylene, —(C₁₋₃ alkylene)_(a)-O—(C₁₋₃alkylene)_(b)-, —(C₁₋₃ alkylene)_(a)-S—(C₁₋₃ alkylene)_(b)-, or —(C₁₋₃alkylene)_(a)-NH—(C₁₋₃ alkylene)_(b)-, wherein the C₁₋₃ alkylene issubstituted with 0 to 3 R¹¹; a is an integer of 0 or 1; b is an integerof 0 or 1; provided that a and b are not both 1; and L² is a covalentbond.

In any one of the preceding embodiments of Formula (I), the E ring is amoiety selected from:

E¹ and E² are independently CR³, CHR³, N, NR³, O or S;

the dashed line is an optional covalent bond; that is, the dashed linedenotes a covalent bond which is either present or absent;

t is 0, 1 or 2; and

each R³ is independently hydrogen, halo, cyano, hydroxyl, amino, C₁₋₆alkyl, alkylamino, haloalkyl, hydroxyalkyl, aminoalkyl, alkoxyalkyl,haloalkoxyalkyl, alkoxy, or haloalkoxy.

In any one of the preceding embodiments of Formula (I),

the

moiety is selected from

wherein the nitrogen atom is attached to L¹.

In any one of the preceding embodiments of Formula (I), Z is phenyl or5- to 10-membered heteroaryl containing 1 to 3 heteroatoms independentlyselected from N, O, and S containing 1 to 3 heteroatoms independentlyselected from N, O, and S, wherein the phenyl and heteroaryl areindependently substituted with 0 to 5 R⁸, wherein R⁸ is the same asdefined above.

In any one of the preceding embodiments of Formula (I), L¹ is a covalentbond.

In any one of the preceding embodiments of Formula (I), —Z—R^(x) is

wherein the Z moiety is further substituted with 0 to 3 R⁸, and R⁸ isthe same as defined above.

In any one of the preceding embodiments of Formula (I), Y is N.

In any one of the preceding embodiments of Formula (I), Y is CH; and L¹is a covalent bond, O, S, NH, —O—(C₁₋₃ alkylene)-, —S—(C₁₋₃ alkylene)-,or —NH—(C₁₋₃ alkylene)-.

In any one of the preceding embodiments of Formula (I), R² is phenyl or6-membered heteroaryl containing 1 to 3 heteroatoms independentlyselected from N, O, and S containing 1 to 3 heteroatoms independentlyselected from N, O, and S, wherein the phenyl or heteroaryl issubstituted with 0 to 3 R¹⁰. In any one of the preceding embodiments ofFormula (I), L² is a covalent bond.

In one embodiment of Formula (I), the compound is represented by Formula(II):

X¹ is C or N;

X² and X³ are each independently CH, N, O, or S;

Z is phenyl or a 5- to 10-membered heteroaryl containing 1 to 3heteroatoms independently selected from N, O, and S containing 1 to 3heteroatoms independently selected from N, O, and S, wherein the phenyland heteroaryl are independently substituted with 0 to 3 R⁸;

R^(X) is —C(O)OR¹³ or —C(O)NH—S(O)₂R^(e);

R^(e) is C₁₋₆ alkyl or C₃₋₆ cycloalkyl;

R¹ is C₁₋₆ alkyl or C₃₋₅ cycloalkyl, wherein the alkyl or cycloalkyl issubstituted with 0 to 3 R⁹;

R² is phenyl or 6-membered heteroaryl containing 1 to 3 heteroatomsindependently selected from N, O, and S containing 1 to 3 heteroatomsindependently selected from N, O, and S, wherein the phenyl orheteroaryl is substituted with 0 to 3 R¹⁰; and

R⁸, R⁹, R¹⁰, and R¹³ are the same as defined above.

In any one of the preceding embodiments of Formula (II), the

moiety is

or

In any one of the preceding embodiments of Formula (II), R² is phenyl orpyridinyl, each of which is independently substituted with 0 to 3 R¹⁰.

In any one of the preceding embodiments of Formula (I),

the

moiety is

In any one of the preceding embodiments of Formula (I), Z is 8- to10-membered bicyclic heteroaryl, wherein the heteroaryl is independentlysubstituted with 0 to 3 R⁸.

In any one of the preceding embodiments of Formula (I), R^(X) is—C(O)OH.

In one embodiment, the present compounds are represented by Formula(III):

or a stereoisomer, a tautomer, or a pharmaceutically acceptable salt orsolvate thereof; wherein:

-   Z is 6-membered monocyclic heteroaryl containing 1 or 2 nitrogen    atoms, or a 9- to 10-membered bicyclic heteroaryl containing 1 or 3    heteroatoms independently selected from N, O, and S, wherein the    monocyclic or bicyclic heteroaryl is independently substituted with    0 to 3 R⁸;-   R² is phenyl or pyridinyl, wherein the phenyl and pyridinyl are each    independently substituted with 0 to 2 R¹⁰;-   R⁸ is each independently halo, cyano, hydroxyl, C₁₋₄ alkyl, C₁₋₄    haloalkyl, C₁₋₄ alkoxy, or C₁₋₄ haloalkoxy;-   R¹⁰ is each independently halo, C₁₋₄ alkyl, C₁₋₄ haloalkyl, C₁₋₄    alkoxy, or C₁₋₄ haloalkoxy;-   R^(X) is —C(O)OH or —C(O)NH—S(O)₂R^(e); and-   R^(e) is C₁₋₆ alkyl or C₃₋₆ cycloalkyl.

In some embodiments of Formula (III), Z is a heteroaryl selected frompyridinyl, benzthiazolyl, pyrrolopyridinyl, pyrrolopyrimidinyl, indolyl,quinolinyl, imidazopyridinyl, pyrazolopyrimidinyl, and pyrrolotriazinyl,wherein the heteroaryl is independently substituted with 0 to 3 R⁸. Insome embodiments, the heteroaryl is independently substituted with 0, 1,or 2 R⁸.

In some embodiments of Formula (III), R² is phenyl or pyridinyl, whereinthe phenyl and pyridinyl are each independently substituted with 1 or 2R¹⁰.

In some embodiments of Formula (III), R⁸ is each independently F, —CH₃,—OCH₃, —OCH₂CH₃, —OCH(CH₃)₂, —CF₃, —OCF₃, or —OCHF₂.

In some embodiments of Formula (III), R¹⁰ is each independently C₁,—CH₃, —CF₃, or —OCF₃.

In some embodiments of Formula (III), R^(X) is —C(O)OH or—C(O)NH—S(O)₂R^(e); and R^(e) is methyl, ethyl, isopropyl, orcyclopropyl.

In one embodiment of Formula (I) or Formula (II), X¹ is C.

In one embodiment of Formula (I) or Formula (II), X² is N.

In one embodiment of Formula (I) or Formula (II), X³ is O.

In one embodiment of Formula (I), X⁴ is C.

In one embodiment of Formula (I), X¹ is C and X⁴ is C.

In one embodiment of Formula (I) or Formula (II), one of X² and X³ is Nand the other of X² and X³ is O.

In one embodiment of Formula (I) or Formula (II), X² is N and X³ is O.

In one embodiment of Formula (I) or Formula (II), X² is O and X³ is N.

In one embodiment of Formula (I) or Formula (II), X¹ is C; X² is N; andX³ is O.

In one embodiment of Formula (I), X¹ is C; one of X² and X³ is N and theother of X² and X³ is O; and X⁴ is C.

In one embodiment of Formula (I), X¹ is C; X² is N; X³ is 0; and X⁴ isC.

In one embodiment of Formula (I), X¹ is C; X² is O; X³ is N; and X⁴ isC.

In one embodiment of Formula (I) or Formula (II), X¹ is N; X² is N; andX³ is N.

In one embodiment of Formula (I), the

moiety is selected from:

In one embodiment of Formula (I), L¹ is a covalent bond, O, —CH₂—,—CH₂CH₂—, —OCH₂—, —CH₂OCH₂—, or —NR¹⁶—. Included in this embodiment arecompounds in which L¹ is a covalent bond, O, or —OCH₂—. Also included inthis embodiment are compounds in which L¹ is a covalent bond.

In one embodiment of Formula (I), the

moiety is selected from:

andL¹ is a covalent bond, O, —CH₂—, —CH₂CH₂—, —OCH₂—, —CH₂OCH₂—, or NR¹⁶—.Included in this embodiment are compounds in which L¹ is a covalentbond, O, or —OCH₂—. Also included in this embodiment are compounds inwhich L¹ is a covalent bond.

In one embodiment of Formula (I), the

moiety is selected from:

andL¹ is a covalent bond, —CH₂—, —CH₂CH₂—, —OCH₂—, —CH₂OCH₂—. Included inthis embodiment are compounds in which L¹ is a covalent bond.

In one embodiment of Formula (I), the

moiety is selected from:

andL¹ is a covalent bond, O, —CH₂—, —CH₂CH₂—, —OCH₂—, —CH₂OCH₂—, or —NR¹⁶—.Included in this embodiment are compounds in which L¹ is a covalentbond, O, or —OCH₂—. Also included in this embodiment are compounds inwhich L¹ is a covalent bond.

In one embodiment of Formula (I), Formula (II), or Formula (III), Z isaryl or 5- to 10-membered heteroaryl containing 1 to 3 heteroatomsindependently selected from N, O, and S, wherein the phenyl andheteroaryl are independently substituted with 0 to 5 R⁸. Included inthis embodiment are compounds in which Z is phenyl, pyridinyl,pyridazinyl, pyrimidinyl, pyrazinyl, benzo[d]imidazolyl,benzo[d]isoxazolyl, benzo[d]oxadiazolyl, benzo[d]thiazolyl,imidazolo[1,5-a]pyridinyl, indazolyl, indolyl, pyrazolo[4,3-b]pyridinyl,pyrrolo[2,1-f][1,2,4]triazinyl, pyrrolo[2,3-b]pyridinyl,pyrrolo[2,3-c]pyridinyl, pyrrolo[2,3-d]pyrimidinyl,pyrrolo[3,2-c]pyridinyl, thiazolo[4,5-b]pyridinyl,thiazolo[5,4-b]pyridinyl, cinnolinyl, isoquinolinyl, quinolinyl, orquinoxalinyl, each substituted with zero to 1 R⁸.

In one embodiment of Formula (I), the

moiety is selected from:

-   L¹ is a covalent bond, O, or —OCH₂—; and-   Z is phenyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl,    benzo[d]imidazolyl, benzo[d]isoxazolyl, benzo[d]oxadiazolyl,    benzo[d]thiazolyl, imidazolo[1,5-a]pyridinyl, indazolyl, indolyl,    pyrazolo[4,3-b]pyridinyl, pyrrolo[2,1-f][1,2,4]triazinyl,    pyrrolo[2,3-b]pyridinyl, pyrrolo[2,3-c]pyridinyl,    pyrrolo[2,3-d]pyrimidinyl, pyrrolo[3,2-c]pyridinyl,    thiazolo[4,5-b]pyridinyl, thiazolo[5,4-b]pyridinyl, cinnolinyl,    isoquinolinyl, quinolinyl, or quinoxalinyl, each substituted with    zero to 1 R⁸.

In one embodiment of Formula (I), Formula (II), or Formula (III), L³ isa covalent bond, —CH₂—, —CH₂CH₂—, —C(O)NHCH₂—, or —OCH₂—.

In one embodiment of Formula (I), Formula (II), or Formula (III), L³ isa covalent bond.

In one embodiment of Formula (I), Formula (II), or Formula (III), L³ is—CH₂—, —CH₂CH₂—, —C(O)NHCH₂—, or —OCH₂—.

In one embodiment of Formula (I), Formula (II), or Formula (III), L³ isa covalent bond or —C(O)NHCH₂—.

In one embodiment of Formula (I), Formula (II), or Formula (III), R^(Z)is —CN, —C(O)OR¹³, —C(O)NR^(14a)R^(14b),

Included in this embodiment are compounds in which L³ is a covalentbond.

In one embodiment of Formula (I), Formula (II), or Formula (III), R^(x)is —CN, —C(O)OH, —C(O)O(C₁₋₃ alkyl), —C(O)NH₂, —C(O)NH(C₁₋₃ alkyl),—C(O)NH(C₃₋₆ cyclopropyl), —C(O)NHCH₂C(O)OH, —C(O)NHS(O)₂(C₁₋₃ alkyl),—C(O)NHS(O)₂(C₃₋₆ cyclopropyl), —OCH₂C(O)OH, or—C(O)O(carboxy(trihydroxy)tetrahydropyranyl). Included in thisembodiment are compounds in which R^(x) is —CN, —C(O)OH, —C(O)OCH₂CH₃,—C(O)NH₂, —C(O)NH(CH₃), —C(O)NHCH₂CH₃, —C(O)NHCH(CH₃)₂,—C(O)NH(cyclopropyl), —C(O)NHCH₂C(O)OH, —C(O)NHS(O)₂CH₃,—C(O)NHS(O)₂(cyclopropyl), —OCH₂C(O)OH, or—C(O)O(carboxy(trihydroxy)tetrahydropyranyl).

In one embodiment of Formula (I), Formula (II), or Formula (III), L² isa covalent bond or —CH(cyclopropyl)-.

In one embodiment of Formula (I), Formula (II), or Formula (III), L² isa covalent bond.

In one embodiment of Formula (I), Formula (II), or Formula (III), L² is—CH(cyclopropyl)- and R² is cyclopropyl.

In one embodiment of Formula (I), Formula (II), or Formula (III), R² isC₃₋₆ cycloalkyl, phenyl, or pyridinyl, wherein the phenyl and thepyridinyl are independently substituted with 1 to 3 R¹⁰.

In one embodiment of Formula (I), Formula (II), or Formula (III), R² iscyclopropyl, cyclohexyl, phenyl, or pyridinyl, wherein the phenyl andthe pyridinyl are independently substituted with 1 to 3 R¹⁰. In oneembodiment of Formula (I), Formula (II), or Formula (III), R² iscyclohexyl, phenyl, or pyridinyl, wherein the phenyl and the pyridinylare independently substituted with 1 to 3 R¹⁰; and L² is a covalentbond.

In one embodiment of Formula (I), Formula (II), or Formula (III), R¹ isC₁₋₃ alkyl, C₃₋₄ cycloalkyl, or C₄₋₅ heterocyclyl, wherein the alkyl,cycloalkyl, and heterocyclyl are each substituted with 0 to 3 R⁹;

In one embodiment of Formula (I), Formula (II), or Formula (III), R¹ is—CHF₂, —CH(CH₃)₂, cyclopropyl, or methylcyclopropyl.

One embodiment provides a compound according to Formula (I) wherein: X¹is C, X² is N, X³ is O, and X⁴ is C; or X¹ is N, X² is N, X³ is C, andX⁴ is C; Y is CH or N;

the

moiety is selected from:

-   L¹ is a covalent bond, O, or —OCH₂—, provided that L¹ is a covalent    bond when Y is N;-   Z is phenyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl,    benzo[d]imidazolyl, benzo[d]isoxazolyl, benzo[d]oxadiazolyl,    benzo[d]thiazolyl, imidazolo[1,5-a]pyridinyl, indazolyl, indolyl,    pyrazolo[4,3-b]pyridinyl, pyrrolo[2,1-f][1,2,4]triazinyl,    pyrrolo[2,3-b]pyridinyl, pyrrolo[2,3-c]pyridinyl,    pyrrolo[2,3-d]pyrimidinyl, pyrrolo[3,2-c]pyridinyl,    thiazolo[4,5-b]pyridinyl, thiazolo[5,4-b]pyridinyl, cinnolinyl,    isoquinolinyl, quinolinyl, or quinoxalinyl, each substituted with    zero to 1 R⁸;-   R⁸ is F, —CH₃, —CF₃, —OCH₃, —OCH₂CH₃, —OCH(CH₃)₂, —OCHF₂, —OCF₃,    —OCH₂CH₂OH, or —CH₂OCH₂CH₂Si(CH₃)₃;-   R^(x) is —CN, —C(O)OH, —C(O)OCH₂CH₃, —C(O)NH₂, —C(O)NH(CH₃),    —C(O)NHCH₂CH₃, —C(O)NHCH(CH₃)₂, —C(O)NH(cyclopropyl),    —C(O)NHCH₂C(O)OH, —C(O)NHS(O)₂CH₃, —C(O)NHS(O)₂(cyclopropyl),    —OCH₂C(O)OH, or —C(O)O(carboxy(trihydroxy)tetrahydropyranyl);-   L² is a covalent bond;-   R¹ is —CHF₂, —CH(CH₃)₂, cyclopropyl, or methylcyclopropyl;-   R² is cyclohexyl, phenyl, or pyridinyl, wherein the phenyl and the    pyridinyl are independently substituted with 1 to 3 R¹⁰; and-   R¹⁰ is each independently F, C₁, —CH₃, —CF₃, —OCH₃, or —OCF₃.

In one embodiment, the present compounds are represented by Formula(III) or a stereoisomer, a tautomer, or a pharmaceutically acceptablesalt or solvate thereof; wherein: Z is a 9- to 10-membered bicyclicheteroaryl containing 1 or 3 heteroatoms independently selected from N,O, and S, wherein the bicyclic heteroaryl is independently substitutedwith 0 to 3 R⁸;

-   R² is phenyl or pyridinyl, wherein the phenyl and pyridinyl are each    independently substituted with 0 to 2 R¹⁰;-   R⁸ is each independently F, C₁, cyano, hydroxyl, C₁₋₃ alkyl, C₁₋₂    haloalkyl, C₁₋₃ alkoxy, or C₁₋₂ haloalkoxy;-   R¹⁰ is each independently F, C₁, C₁₋₃ alkyl, C₁₋₂ fluoroalkyl, C₁₋₄    alkoxy, or C₁₋₂ fluoroalkoxy;-   R^(X) is —C(O)OH.

In one embodiment, the present compounds are represented by Formula(III) or a stereoisomer, a tautomer, or a pharmaceutically acceptablesalt or solvate thereof; wherein:

-   Z is a 9-membered bicyclic heteroaryl containing 1 or 3 heteroatoms    independently selected from N, O, and S, wherein the bicyclic    heteroaryl is independently substituted with 0 to 3 R⁸;-   R² is phenyl or pyridinyl, wherein the phenyl and pyridinyl are each    independently substituted with 0 to 2 R¹⁰;-   R⁸ is each independently F, C₁, cyano, hydroxyl, C₁₋₃ alkyl, C₁₋₂    haloalkyl, C₁₋₃ alkoxy, or C₁₋₂ haloalkoxy;-   R¹⁰ is each independently F, C₁, C₁₋₃ alkyl, C₁₋₂ fluoroalkyl, C₁₋₄    alkoxy, or C₁₋₂ fluoroalkoxy; and-   R^(X) is —C(O)OH.

In one embodiment, the present invention provides compounds selectedfrom:

In one embodiment, the present invention provides compounds selectedfrom:

In one embodiment, the present invention provides a compound selectedfrom:

or a stereoisomer, a tautomer, or a pharmaceutically acceptable salt orsolvate thereof.

In one embodiment, the present invention provides, inter alia, compoundsselected from any one of the Examples as described in the specification,or a stereoisomer, a tautomer, or a pharmaceutically acceptable salt orsolvate thereof.

In one embodiment, the compounds of the present invention have FXR EC₅₀values ≤5000 nM, using the transient human FXR/Gal4-luciferase reporterassay; in another embodiment, the compounds of the present inventionhave FXR EC₅₀ values ≤1000 nM; in another embodiment, the compounds ofthe present invention have FXR EC₅₀ values ≤500 nM; in anotherembodiment, the compounds of the present invention have FXR EC₅₀ values≤200 nM; in another embodiment, the compounds of the present inventionhave FXR EC₅₀ values ≤100 nM; in another embodiment, the compounds ofthe present invention have FXR EC₅₀ values ≤50 nM.

II. Pharmaceutical Compositions, Therapeutic Utilities, and Combinations

In another embodiment, the present invention provides a compositioncomprising at least one of the compounds of the present invention, or astereoisomer, a tautomer, or a pharmaceutically acceptable salt or asolvate thereof.

In another embodiment, the present invention provides a pharmaceuticalcomposition comprising a pharmaceutically acceptable carrier and atleast one of the compounds of the present invention or a stereoisomer, atautomer, or a pharmaceutically acceptable salt or a solvate thereof.

In another embodiment, the present invention provides a pharmaceuticalcomposition, comprising a pharmaceutically acceptable carrier and atherapeutically effective amount of at least one of the compounds of thepresent invention or a stereoisomer, a tautomer, or a pharmaceuticallyacceptable salt or a solvate thereof.

In another embodiment, the present invention provides a process formaking a compound of the present invention.

In another embodiment, the present invention provides an intermediatefor making a compound of the present invention.

In another embodiment, the present invention provides a pharmaceuticalcomposition as defined above further comprising one or more additionaltherapeutic agents.

In another embodiment, the present invention provides a method for thetreatment of a disease, disorder, or condition associated withdysregulation of bile acids in a patient in need of such treatment, andthe method comprises administering a therapeutically effective amount ofa compound of the present invention, or a stereoisomer, a tautomer, or apharmaceutically acceptable salt or solvate thereof, to the patient.

In another embodiment, the present invention provides a method for thetreatment of a disease, disorder, or condition associated with activityof famesoid X receptor (FXR) in a patient in need of such treatmentcomprising administering a therapeutically effective amount of acompound of the present invention, or a stereoisomer, a tautomer, or apharmaceutically acceptable salt or solvate thereof, to the patient.

In another embodiment, the present invention provides a method for thetreatment of the disease, disorder, or condition comprisingadministering to a patient in need of such treatment a therapeuticallyeffective amount of at least one of the compounds of the presentinvention, alone, or, optionally, in combination with another compoundof the present invention and/or at least one other type of therapeuticagent.

In another embodiment, the present invention provides a method foreliciting an famesoid X receptor (FXR) agonizing effect in a patientcomprising administering a therapeutically effective amount of acompound of the present invention, or a stereoisomer, a tautomer, or apharmaceutically acceptable salt or solvate thereof, to the patient.

In some embodiments, the disease, disorder, or condition is associatedwith FXR dysfunction include pathological fibrosis, cancer, inflammatorydisorders, metabolic, or cholestatic disorders.

In some embodiments, the disease, disorder, or condition is associatedwith fibrosis, including liver, biliary, renal, cardiac, dermal, ocular,and pancreatic fibrosis.

In other embodiments, the disease, disorder, or condition is associatedwith cell-proliferative disorders, such as cancer. In some embodiments,the cancer includes solid tumor growth or neoplasia. In otherembodiments, the cancer includes tumor metastasis. In some embodiments,the cancer is of the liver, gall bladder, small intestine, largeintestine, kidney, prostate, bladder, blood, bone, brain, breast,central nervous system, cervix, colon, endometrium, esophagus,genitalia, genitourinary tract, head, larynx, lung, muscle tissue, neck,oral or nasal mucosa, ovary, pancreas, skin, spleen, stomach, testicle,or thyroid. In other embodiments, the cancer is a carcinoma, sarcoma,lymphoma, leukemia, melanoma, mesothelioma, multiple myeloma, orseminoma.

Examples of diseases, disorders, or conditions associated with theactivity of FXR that can be prevented, modulated, or treated accordingto the present invention include, but are not limited to, transplantinjection, fibrotic disorders (e.g., liver fibrosis, kidney fibrosis),inflammatory disorders (e.g., acute hepatitis, chronic hepatitis,non-alcoholic steatohepatitis (NASH), irritable bowel syndrome (IBS),inflammatory bowel disease (IBD)), as well as cell-proliferativedisorders (e.g., cancer, myeloma, fibroma, hepatocellular carcinoma,colorectal cancer, prostate cancer, leukemia, Kaposi's sarcoma, solidtumors).

The fibrotic disorders, inflammatory disorders, as well ascell-proliferative disorders that are suitable to be prevented ortreated by the compounds of the present invention include, but are notlimited to, non-alcoholic fatty liver disease (NAFLD), alcoholic ornon-alcoholic steatohepatitis (NASH), acute hepatitis, chronichepatitis, liver cirrhosis, primary biliary cirrhosis, primarysclerosing cholangitis, drug-induced hepatitis, biliary cirrhosis,portal hypertension, regenerative failure, liver hypofunction, hepaticblood flow disorder, nephropathy, irritable bowel syndrome (IBS),inflammatory bowel disease (IBD), abnormal pancreatic secretion, benignprostatic hyperplasia, neuropathic bladder disease, diabeticnephropathy, focal segmental glomerulosclerosis, IgA nephropathy,nephropathy induced by drugs or transplantation, autoimmune nephropathy,lupus nephritis, liver fibrosis, kidney fibrosis, chronic kidney disease(CKD), diabetic kidney disease (DKD), skin fibrosis, keloids, systemicsclerosis, scleroderma, virally-induced fibrosis, idiopathic pulmonaryfibrosis (IPF), interstitial lung disease, non-specific interstitialpneumonia (NSIP), usual interstitial pneumonia (UIP), radiation-inducedfibrosis, familial pulmonary fibrosis, airway fibrosis, chronicobstructive pulmonary disease (COPD), spinal cord tumor, hernia ofintervertebral disk, spinal canal stenosis, heart failure, cardiacfibrosis, vascular fibrosis, perivascular fibrosis, foot-and-mouthdisease, cancer, myeloma, fibroma, hepatocellular carcinoma, colorectalcancer, prostate cancer, leukemia, chronic lymphocytic leukemia,Kaposi's sarcoma, solid tumors, cerebral infarction, cerebralhemorrhage, neuropathic pain, peripheral neuropathy, age-related maculardegeneration (AMD), glaucoma, ocular fibrosis, corneal scarring,diabetic retinopathy, proliferative vitreoretinopathy (PVR), cicatricialpemphigoid glaucoma filtration surgery scarring, Crohn's disease orsystemic lupus erythematosus; keloid formation resulting from abnormalwound healing; fibrosis occurring after organ transplantation,myelofibrosis, and fibroids. In one embodiment, the present inventionprovides a method for the treatment of a fibrotic disorder, aninflammatory disorder, or a cell-proliferative disorder, comprisingadministering to a patient in need of such treatment a therapeuticallyeffective amount of at least one of the compounds of the presentinvention, alone, or, optionally, in combination with another compoundof the present invention and/or at least one other type of therapeuticagent.

In another embodiment, the present invention provides a compound of thepresent invention for use in therapy.

In another embodiment, the present invention provides a compound of thepresent invention for use in therapy for the treatment of a fibroticdisorder, an inflammatory disorder, or a cell-proliferative disorderthereof.

In another embodiment, the present invention also provides the use of acompound of the present invention for the manufacture of a medicamentfor the treatment of a fibrotic disorder, an inflammatory disorder, or acell-proliferative disorder thereof.

In another embodiment, the present invention provides a method for thetreatment of a fibrotic disorder, an inflammatory disorder, or acell-proliferative disorder, comprising administering to a patient inneed thereof a therapeutically effective amount of a first and secondtherapeutic agent, wherein the first therapeutic agent is a compound ofthe present invention.

In another embodiment, the present invention provides a combinedpreparation of a compound of the present invention and additionaltherapeutic agent(s) for simultaneous, separate or sequential use intherapy.

In another embodiment, the present invention provides a combinedpreparation of a compound of the present invention and additionaltherapeutic agent(s) for simultaneous, separate or sequential use in thetreatment of a fibrotic disorder, an inflammatory disorder, or acell-proliferative disorder.

The compounds of the present invention may be employed in combinationwith additional therapeutic agent(s), such as one or more anti-fibroticand/or anti-inflammatory therapeutic agents.

In one embodiment, additional therapeutic agent(s) used in combinedpharmaceutical compositions or combined methods or combined uses, areselected from one or more, preferably one to three, of the followingtherapeutic agents: TGFβ receptor inhibitors (for example,galunisertib), inhibitors of TGFβ synthesis (for example, pirfenidone),inhibitors of vascular endothelial growth factor (VEGF),platelet-derived growth factor (PDGF) and fibroblast growth factor (FGF)receptor kinases (for example, nintedanib), humanized anti-αvβ6 integrinmonoclonal antibody (for example, 3G9), human recombinant pentraxin-2,recombinant human Serum Amyloid P, recombinant human antibody againstTGFβ-1, -2, and -3, endothelin receptor antagonists (for example,macitentan), interferon gamma, c-Jun amino-terminal kinase (JNK)inhibitor (for example,4-[[9-[(3S)-tetrahydro-3-furanyl]-8-[(2,4,6-trifluorophenyl)amino]-9H-purin-2-yl]amino]-trans-cyclohexanol,3-pentylbenzeneacetic acid (PBI-4050), tetra-substituted porphyrinderivative containing manganese (III), monoclonal antibody targetingeotaxin-2, interleukin-13 (IL-13) antibody (for example, lebrikizumab,tralokinumab), bispecific antibody targeting interleukin 4 (IL-4) andinterleukin 13 (IL-13), NK1 tachykinin receptor agonist (for example,Sar⁹, Met(O₂)¹¹-Substance P), Cintredekin Besudotox, human recombinantDNA-derived, IgG1 kappa monoclonal antibody to connective growth factor,and fully human IgG1 kappa antibody, selective for CC-chemokine ligand 2(for example, carlumab, CCX140), antioxidants (for example,N-acetylcysteine), phosphodiesterase 5 (PDE5) inhibitors (for example,sildenafil), agents for treatment of obstructive airway diseases such asmuscarinic antagonists (for example, tiotropium, ipatropium bromide),adrenergic β2 agonists (for example, salbutamol, salmeterol),corticosteroids (for example, triamcinolone, dexamethasone,fluticasone), immunosuppressive agents (for example, tacrolimus,rapamycin, pimecrolimus), and therapeutic agents useful for thetreatment of fibrotic conditions, such as liver, biliary, and kidneyfibrosis, Non-Alcoholic Fatty Liver Disease (NALFD), Non-AlcoholicSteato-Hepatitis (NASH), cardiac fibrosis, Idiopathic Pulmonary Fibrosis(IPF), and systemic sclerosis. The therapeutic agents useful for thetreatment of such fibrotic conditions include, but are not limited to,FXR agonists (for example OCA, GS-9674, and LJN452), LOXL2 inhibitors(for example simtuzumab), LPA1 antagonists (for example, BMS-986020 andSAR 100842), PPAR modulators (for example, elafibrinor, pioglitazone,and saroglitazar, IVA337), SSAO/VAP-1 inhibitors (for example, PXS-4728Aand SZE5302), ASK-1 inhibitors (for example GS-4997 or selonsertib), ACCinhibitors (for example, CP-640186 and NDI-010976 or GS-0976), FGF21mimetics (for example, LY2405319 and BMS-986036), caspase inhibitors(for example, emricasan), NOX4 inhibitors (for example, GKT137831),MGAT2 inhibitor (for example, BMS-963272), αV integrin inhibitors (forexample, abituzumab) and bile acid/fatty acid conjugates (for examplearamchol). The FXR agonists of various embodiments of the presentinvention may also be used in combination with one or more therapeuticagents such as CCR2/5 inhibitors (for example, cenicriviroc), Galectin-3inhibitors (for example, TD-139, GR-MD-02), leukotriene receptorantagonists (for example, tipelukast, montelukast), SGLT2 inhibitors(for example, dapagliflozin, remogliflozin), GLP-1 receptor agonists(for example, liraglutide and semaglutide), FAK inhibitors (for example,GSK-2256098), CB1 inverse agonists (for example, JD-5037), CB2 agonists(for example, APD-371 and JBT-101), autotaxin inhibitors (for example,GLPG1690), prolyl t-RNA synthetase inhibitors (for example,halofugenone), FPR2 agonists (for example, ZK-994), and THR agonists(for example, MGL:3196). In another embodiment, additional therapeuticagent(s) used in combined pharmaceutical compositions or combinedmethods or combined uses, are selected from one or more, preferably oneto three, of immunoncology agents, such as Alemtuzumab, Atezolizumab,Ipilimumab, Nivolumab, Ofatumumab, Pembrolizumab, and Rituximab.

The compounds of this invention can be administered for any of the usesdescribed herein by any suitable means, for example, orally, such astablets, capsules (each of which includes sustained release or timedrelease formulations), pills, powders, granules, elixirs, tinctures,suspensions, syrups, and emulsions; sublingually; bucally; parenterally,such as by subcutaneous, intravenous, intramuscular, or intrastemalinjection, or infusion techniques (e.g., as sterile injectable aqueousor non-aqueous solutions or suspensions); nasally, includingadministration to the nasal membranes, such as by inhalation spray;topically, such as in the form of a cream or ointment; or rectally suchas in the form of suppositories. They can be administered alone, butgenerally will be administered with a pharmaceutical carrier selected onthe basis of the chosen route of administration and standardpharmaceutical practice.

The term “pharmaceutical composition” means a composition comprising acompound of the invention in combination with at least one additionalpharmaceutically acceptable carrier. A “pharmaceutically acceptablecarrier” refers to media generally accepted in the art for the deliveryof biologically active agents to animals, in particular, mammals,including, i.e., adjuvant, excipient or vehicle, such as diluents,preserving agents, fillers, flow regulating agents, disintegratingagents, wetting agents, emulsifying agents, suspending agents,sweetening agents, flavoring agents, perfuming agents, anti-bacterialagents, anti-fungal agents, lubricating agents and dispensing agents,depending on the nature of the mode of administration and dosage forms.Pharmaceutically acceptable carriers are formulated according to anumber of factors well within the purview of those of ordinary skill inthe art. These include, without limitation: the type and nature of theactive agent being formulated; the subject to which the agent-containingcomposition is to be administered; the intended route of administrationof the composition; and the therapeutic indication being targeted.Pharmaceutically acceptable carriers include both aqueous and nonaqueousliquid media, as well as a variety of solid and semi-solid dosage forms.Such carriers can include a number of different ingredients andadditives in addition to the active agent, such additional ingredientsbeing included in the formulation for a variety of reasons, e.g.,stabilization of the active agent, binders, etc., well known to those ofordinary skill in the art. Descriptions of suitable pharmaceuticallyacceptable carriers, and factors involved in their selection, are foundin a variety of readily available sources such as, for example,Remington's Pharmaceutical Sciences, 18th Edition (1990).

The terms “treating” or “treatment” as used herein refer to an approachfor obtaining beneficial or desired results, including clinical results,by using a compound or a composition of the present invention. Forpurposes of this invention, beneficial or desired clinical resultsinclude, but are not limited to, one or more of the following:decreasing the severity and/or frequency one or more symptoms resultingfrom the disease, disorder, or condition; diminishing the extent of orcausing regression of the disease, disorder, or condition; stabilizingthe disease, disorder, or condition (e.g., preventing or delaying theworsening of the disease, disorder, or condition); delay or slowing theprogression of the disease, disorder, or condition; ameliorating thedisease, disorder, or condition state; decreasing the dose of one ormore other medications required to treat the disease, disorder, orcondition; and/or increasing the quality of life.

The dosage regimen for the compounds of the present invention will, ofcourse, vary depending upon known factors, such as the pharmacodynamiccharacteristics of the particular agent and its mode and route ofadministration; the species, age, sex, health, medical condition, andweight of the recipient; the nature and extent of the symptoms; the kindof concurrent treatment; the frequency of treatment; the route ofadministration, the renal and hepatic function of the patient, and theeffect desired.

By way of general guidance, the daily oral dosage of each activeingredient, when used for the indicated effects, will range betweenabout 0.01 to about 5000 mg per day, preferably between about 0.01 toabout 1000 mg per day, and most preferably between about 0.01 to about250 mg per day. Intravenously, the most preferred doses will range fromabout 0.01 to about 10 mg/kg/minute during a constant rate infusion.Compounds of this invention may be administered in a single daily dose,or the total daily dosage may be administered in divided doses of two,three, or four times daily.

The compounds are typically administered in admixture with suitablepharmaceutical diluents, excipients, or carriers (collectively referredto herein as pharmaceutical carriers) suitably selected with respect tothe intended form of administration, e.g., oral tablets, capsules,elixirs, and syrups, and consistent with conventional pharmaceuticalpractices.

Dosage forms (pharmaceutical compositions) suitable for administrationmay contain from about 0.1 milligram to about 2000 milligrams of activeingredient per dosage unit. In these pharmaceutical compositions theactive ingredient will ordinarily be present in an amount of about0.1-95% by weight based on the total weight of the composition.

A typical capsule for oral administration contains at least one of thecompounds of the present invention (250 mg), lactose (75 mg), andmagnesium stearate (15 mg). The mixture is passed through a 60 meshsieve and packed into a No. 1 gelatin capsule.

A typical injectable preparation is produced by aseptically placing atleast one of the compounds of the present invention (250 mg) into avial, aseptically freeze-drying and sealing. For use, the contents ofthe vial are mixed with 2 mL of physiological saline, to produce aninjectable preparation.

The present invention includes within its scope pharmaceuticalcompositions comprising, as an active ingredient, a therapeuticallyeffective amount of at least one of the compounds of the presentinvention, alone or in combination with a pharmaceutical carrier.Optionally, compounds of the present invention can be used alone, incombination with other compounds of the invention, or in combinationwith one or more, preferably one to three, other therapeutic agent(s),e.g., ASK-1 inhibitors, CCR2/5 antagonists, autotaxin inhibitors, LPA1receptor antagonists or other pharmaceutically active material.

The above other therapeutic agents, when employed in combination withthe compounds of the present invention may be used, for example, inthose amounts indicated in the Physicians' Desk Reference, as in thepatents set out above, or as otherwise determined by one of ordinaryskill in the art.

Particularly when provided as a single dosage unit, the potential existsfor a chemical interaction between the combined active ingredients. Forthis reason, when the compound of the present invention and a secondtherapeutic agent are combined in a single dosage unit they areformulated such that although the active ingredients are combined in asingle dosage unit, the physical contact between the active ingredientsis minimized (that is, reduced). For example, one active ingredient maybe enteric coated. By enteric coating one of the active ingredients, itis possible not only to minimize the contact between the combined activeingredients, but also, it is possible to control the release of one ofthese components in the gastrointestinal tract such that one of thesecomponents is not released in the stomach but rather is released in theintestines. One of the active ingredients may also be coated with amaterial that affects a sustained-release throughout thegastrointestinal tract and also serves to minimize physical contactbetween the combined active ingredients. Furthermore, thesustained-released component can be additionally enteric coated suchthat the release of this component occurs only in the intestine. Stillanother approach would involve the formulation of a combination productin which the one component is coated with a sustained and/or entericrelease polymer, and the other component is also coated with a polymersuch as a low viscosity grade of hydroxypropyl methylcellulose (HPMC) orother appropriate materials as known in the art, in order to furtherseparate the active components. The polymer coating serves to form anadditional barrier to interaction with the other component.

These as well as other ways of minimizing contact between the componentsof combination products of the present invention, whether administeredin a single dosage form or administered in separate forms but at thesame time by the same manner, will be readily apparent to those skilledin the art, once armed with the present disclosure.

The compounds of the present invention can be administered alone or incombination with one or more, preferably one to three, additionaltherapeutic agents. By “administered in combination” or “combinationtherapy” it is meant that the compound of the present invention and oneor more, preferably one to three, additional therapeutic agents areadministered concurrently to the mammal being treated. When administeredin combination, each component may be administered at the same time orsequentially in any order at different points in time. Thus, eachcomponent may be administered separately but sufficiently closely intime so as to provide the desired therapeutic effect.

The compounds of the present invention are also useful as standard orreference compounds, for example as a quality standard or control, intests or assays involving FXR agonists. Such compounds may be providedin a commercial kit, for example, for use in pharmaceutical researchinvolving FXR agonist activity. For example, a compound of the presentinvention could be used as a reference in an assay to compare its knownactivity to a compound with an unknown activity. This would ensure theexperimenter that the assay was being performed properly and provide abasis for comparison, especially if the test compound was a derivativeof the reference compound. When developing new assays or protocols,compounds according to the present invention could be used to test theireffectiveness.

The present invention also encompasses an article of manufacture. Asused herein, article of manufacture is intended to include, but not belimited to, kits and packages. The article of manufacture of the presentinvention, comprises: (a) a first container; (b) a pharmaceuticalcomposition located within the first container, wherein the composition,comprises: a first therapeutic agent, comprising a compound of thepresent invention or a pharmaceutically acceptable salt form thereof;and, (c) a package insert stating that the pharmaceutical compositioncan be used for the treatment of dyslipidemias and the sequelae thereof.In another embodiment, the package insert states that the pharmaceuticalcomposition can be used in combination (as defined previously) with asecond therapeutic agent for the treatment of fibrosis and the sequelaethereof. The article of manufacture can further comprise: (d) a secondcontainer, wherein components (a) and (b) are located within the secondcontainer and component (c) is located within or outside of the secondcontainer. Located within the first and second containers means that therespective container holds the item within its boundaries.

The first container is a receptacle used to hold a pharmaceuticalcomposition. This container can be for manufacturing, storing, shipping,and/or individual/bulk selling. First container is intended to cover abottle, jar, vial, flask, syringe, tube (e.g., for a cream preparation),or any other container used to manufacture, hold, store, or distribute apharmaceutical product.

The second container is one used to hold the first container and,optionally, the package insert. Examples of the second containerinclude, but are not limited to, boxes (e.g., cardboard or plastic),crates, cartons, bags (e.g., paper or plastic bags), pouches, and sacks.The package insert can be physically attached to the outside of thefirst container via tape, glue, staple, or another method of attachment,or it can rest inside the second container without any physical means ofattachment to the first container. Alternatively, the package insert islocated on the outside of the second container. When located on theoutside of the second container, it is preferable that the packageinsert is physically attached via tape, glue, staple, or another methodof attachment. Alternatively, it can be adjacent to or touching theoutside of the second container without being physically attached.

The package insert is a label, tag, marker, etc. that recitesinformation relating to the pharmaceutical composition located withinthe first container. The information recited will usually be determinedby the regulatory agency governing the area in which the article ofmanufacture is to be sold (e.g., the United States Food and DrugAdministration). Preferably, the package insert specifically recites theindications for which the pharmaceutical composition has been approved.The package insert may be made of any material on which a person canread information contained therein or thereon. Preferably, the packageinsert is a printable material (e.g., paper, plastic, cardboard, foil,adhesive-backed paper or plastic, etc.) on which the desired informationhas been formed (e.g., printed or applied).

III. Definitions

Throughout the specification and the appended claims, a given chemicalformula or name shall encompass all stereo and optical isomers andracemates thereof where such isomers exist. Unless otherwise indicated,all chiral (enantiomeric and diastereomeric) and racemic forms arewithin the scope of the invention. Many geometric isomers of C═C doublebonds, C═N double bonds, ring systems, and the like can also be presentin the compounds, and all such stable isomers are contemplated in thepresent invention. Cis- and trans- (or E- and Z-) geometric isomers ofthe compounds of the present invention are described and may be isolatedas a mixture of isomers or as separated isomeric forms. The presentcompounds can be isolated in optically active or racemic forms.Optically active forms may be prepared by resolution of racemic forms orby synthesis from optically active starting materials. All processesused to prepare compounds of the present invention and intermediatesmade therein are considered to be part of the present invention. Whenenantiomeric or diastereomeric products are prepared, they may beseparated by conventional methods, for example, by chromatography orfractional crystallization. Depending on the process conditions the endproducts of the present invention are obtained either in free (neutral)or salt form. Both the free form and the salts of these end products arewithin the scope of the invention. If so desired, one form of a compoundmay be converted into another form. A free base or acid may be convertedinto a salt; a salt may be converted into the free compound or anothersalt; a mixture of isomeric compounds of the present invention may beseparated into the individual isomers. Compounds of the presentinvention, free form and salts thereof, may exist in multiple tautomericforms, in which hydrogen atoms are transposed to other parts of themolecules and the chemical bonds between the atoms of the molecules areconsequently rearranged. It should be understood that all tautomericforms, insofar as they may exist, are included within the invention. Asused herein, “a compound of the invention” or “compounds of theinvention” means one or more compounds encompassed by any one of Formula(I), (IIa), and (IIb), or stereoisomers, tautomers, or pharmaceuticallyacceptable salts or solvates thereof.

As used herein, the term “alkyl” or “alkylene” is intended to includeboth branched and straight-chain saturated aliphatic hydrocarbon groupshaving the specified number of carbon atoms. While “alkyl” denotes amonovalent saturated aliphatic radical (such as ethyl), “alkylene”denotes a bivalent saturated aliphatic radical (such as ethylene). Forexample, “C₁ to C₁₀ alkyl” or “C₁₋₁₀ alkyl” is intended to include C₁,C₂, C₃, C₄, C₅, C₆, C₇, C₈, C₉, and C₁₀ alkyl groups. “C₁ to C₁₀alkylene” or “C₁₋₁₀ alkylene”, is intended to include C₁, C₂, C₃, C₄,C₅, C₆, C₇, C₈, C₉, and C₁₀ alkylene groups. Additionally, for example,“C₁ to C₆ alkyl” or “C₁₋₆ alkyl” denotes alkyl having 1 to 6 carbonatoms; and “C₁ to C₆ alkylene” or “C₁₋₆ alkylene” denotes alkylenehaving 1 to 6 carbon atoms. Alkyl group can be unsubstituted orsubstituted with at least one hydrogen being replaced by anotherchemical group. Example alkyl groups include, but are not limited to,methyl (Me), ethyl (Et), propyl (e.g., n-propyl and isopropyl), butyl(e.g., n-butyl, isobutyl, t-butyl), and pentyl (e.g., n-pentyl,isopentyl, neopentyl). When “C₀ alkyl” or “C₀ alkylene” is used, it isintended to denote a direct bond.

Unless otherwise indicated, the term “lower alkyl” as employed hereinalone or as part of another group includes both straight and branchedchain hydrocarbons containing 1 to 8 carbons, and the terms “alkyl” and“alk” as employed herein alone or as part of another group includes bothstraight and branched chain hydrocarbons containing 1 to 20 carbons,preferably 1 to 10 carbons, more preferably 1 to 8 carbons, in thenormal chain, such as methyl, ethyl, propyl, isopropyl, butyl, t-butyl,isobutyl, pentyl, hexyl, isohexyl, heptyl, 4,4-dimethylpentyl, octyl,2,2,4-trimethylpentyl, nonyl, decyl, undecyl, dodecyl, the variousbranched chain isomers thereof, and the like.

“Heteroalkyl” refers to an alkyl group where one or more carbon atomshave been replaced with a heteroatom, such as, O, N, or S. For example,if the carbon atom of the alkyl group which is attached to the parentmolecule is replaced with a heteroatom (e.g., O, N, or S) the resultingheteroalkyl groups are, respectively, an alkoxy group (e.g., —OCH₃,etc.), an alkylamino (e.g., —NHCH₃, —N(CH₃)₂, etc.), or a thioalkylgroup (e.g., —SCH₃). If a non-terminal carbon atom of the alkyl groupwhich is not attached to the parent molecule is replaced with aheteroatom (e.g., O, N, or S) and the resulting heteroalkyl groups are,respectively, an alkyl ether (e.g., —CH₂CH₂—O—CH₃, etc.), analkylaminoalkyl (e.g., —CH₂NHCH₃, —CH₂N(CH₃)₂, etc.), or a thioalkylether (e.g., —CH₂—S—CH₃). If a terminal carbon atom of the alkyl groupis replaced with a heteroatom (e.g., O, N, or S), the resultingheteroalkyl groups are, respectively, a hydroxyalkyl group (e.g.,—CH₂CH₂—OH), an aminoalkyl group (e.g., —CH₂NH₂), or an alkyl thiolgroup (e.g., —CH₂CH₂—SH). A heteroalkyl group can have, for example, 1to 20 carbon atoms, 1 to 10 carbon atoms, or 1 to 6 carbon atoms. AC₁-C₆ heteroalkyl group means a heteroalkyl group having 1 to 6 carbonatoms.

“Alkenyl” or “alkenylene” is intended to include hydrocarbon chains ofeither straight or branched configuration having the specified number ofcarbon atoms and one or more, preferably one to two, carbon-carbondouble bonds that may occur in any stable point along the chain. While“alkenyl” denotes a monovalent radical, “alkenylene” denotes a bivalentradical. For example, “C₂ to C₆ alkenyl” or “C₂₋₆ alkenyl” (oralkenylene), is intended to include C₂, C₃, C₄, C₅, and C₆ alkenylgroups. Examples of alkenyl include, but are not limited to, ethenyl,1-propenyl, 2-propenyl, 2-butenyl, 3-butenyl, 2-pentenyl, 3, pentenyl,4-pentenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl,2-methyl-2-propenyl, and 4-methyl-3-pentenyl.

“Alkynyl” or “alkynylene” is intended to include hydrocarbon chains ofeither straight or branched configuration having one or more, preferablyone to three, carbon-carbon triple bonds that may occur in any stablepoint along the chain. While “alkynyl” denotes a monovalent radical,“alkynylene” denotes a bivalent radical. For example, “C₂ to C₆ alkynyl”or “C₂₋₆ alkynyl” (or alkynylene), is intended to include C₂, C₃, C₄,C₅, and C₆ alkynyl groups; such as ethynyl, propynyl, butynyl, pentynyl,and hexynyl.

As used herein, “arylalkyl” (a.k.a. aralkyl), “heteroarylalkyl”“carbocyclylalkyl” or “heterocyclylalkyl” refers to an acyclic alkylradical in which one of the hydrogen atoms bonded to a carbon atom,typically a terminal or sp³ carbon atom, is replaced with an aryl,heteroaryl, carbocyclyl, or heterocyclyl radical, respectively. Typicalarylalkyl groups include, but are not limited to, benzyl,2-phenylethan-1-yl, naphthylmethyl, 2-naphthylethan-1-yl, naphthobenzyl,2-naphthophenylethan-1-yl and the like. The arylalkyl, heteroarylalkyl,carbocyclylalkyl, or heterocyclylalkyl group can comprise 4 to 20 carbonatoms and 0 to 5 heteroatoms, e.g., the alkyl moiety may contain 1 to 6carbon atoms.

The term “benzyl”, as used herein, refers to a methyl group on which oneof the hydrogen atoms is replaced by a phenyl group, wherein said phenylgroup may optionally be substituted with 1 to 5 groups, preferably 1 to3 groups, —OH, —OCH₃, C₁, F, Br, I, —CN, —NO₂, —NH₂, —NH(CH₃), —N(CH₃)₂,—CF₃, —OCF₃, —C(═O)CH₃, —SCH₃, —S(═O)CH₃, —S(═O)₂CH₃, —CH₃, —CH₂CH₃,—CO₂H, and —CO₂CH₃. “Benzyl” can also be represented by formula “Bn”.

The term “lower alkoxy”, “alkoxy” or “alkyloxy”, “aryloxy” or “aralkoxy”refers to any of the above alkyl, aralkyl or aryl groups linked to anoxygen atom. “C₁ to C₆ alkoxy” or “C₁₋₆ alkoxy” (or alkyloxy), isintended to include C₁, C₂, C₃, C₄, C₅, and C₆ alkoxy groups. Examplealkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy(e.g., n-propoxy and isopropoxy), and t-butoxy. Similarly, “loweralkylthio”, “alkylthio”, “thioalkoxy”, “arylthio”, or “aralkylthio”represents an alkyl, aryl, or aralkyl group as defined above with theindicated number of carbon atoms attached through a sulphur bridge; forexample methyl-S- and ethyl-S—.

The term “alkanoyl” or “alkylcarbonyl” as used herein alone or as partof another group refers to alkyl linked to a carbonyl group. Forexample, alkylcarbonyl may be represented by alkyl-C(O)—. “C₁ to C₆alkylcarbonyl” (or alkylcarbonyl), is intended to include C₁, C₂, C₃,C₄, C₅, and C₆ alkyl-C(O)— groups.

The term “alkylsulfonyl” or “sulfonamide” as used herein alone or aspart of another group refers to alkyl or amino linked to a sulfonylgroup. For example, alkylsulfonyl may be represented by —S(O)₂R′, whilesulfonamide may be represented by —S(O)₂NR^(c)R^(d). R′ is C₁ to C₆alkyl; and R and R^(d) are the same as defined below for “amino”.

The term “carbamate” as used herein alone or as part of another grouprefers to oxygen linked to an amido group. For example, carbamate may berepresented by N(R^(c)R^(d))—C(O)—O—, and R^(c) and R^(d) are the sameas defined below for “amino”.

The term “amido” as used herein alone or as part of another group refersto amino linked to a carbonyl group. For example, amido may berepresented by N(R^(c)R^(d))—C(O)—, and R^(c) and R^(d) are the same asdefined below for “amino”.

The term “amino” is defined as —NR^(c1)R^(c2), wherein R^(c1) and R^(c2)are independently H or C₁₋₆ alkyl; or alternatively, R^(c1) and R^(c2),taken together with the atoms to which they are attached, form a 3- to8-membered heterocyclic ring which is optionally substituted with one ormore group selected from halo, cyano, hydroxyl, amino, oxo, C₁₋₆ alkyl,alkoxy, and aminoalkyl. When R^(c1) or R^(c2) (or both of them) is C₁₋₆alkyl, the amino group can also be referred to as alkylamino. Examplesof alkylamino group include, without limitation, —NH₂, methylamino,ethylamino, propylamino, isopropylamino and the like.

The term “aminoalkyl” refers to an alkyl group on which one of thehydrogen atoms is replaced by an amino group. For example, aminoalkylmay be represented by N(R^(c1)R^(c2))-alkylene-. “C₁ to C₆” or “C₁₋₆”aminoalkyl” (or aminoalkyl), is intended to include C₁, C₂, C₃, C₄, C₅,and C₆ aminoalkyl groups.

The term “halogen” or “halo” as used herein alone or as part of anothergroup refers to chlorine, bromine, fluorine, and iodine, with chlorineor fluorine being preferred.

“Haloalkyl” is intended to include both branched and straight-chainsaturated aliphatic hydrocarbon groups having the specified number ofcarbon atoms, substituted with one or more halogens. “C₁ to C₆haloalkyl” or “C₁₋₆ haloalkyl” (or haloalkyl), is intended to includeC₁, C₂, C₃, C₄, C₅, and C₆ haloalkyl groups. Examples of haloalkylinclude, but are not limited to, fluoromethyl, difluoromethyl,trifluoromethyl, trichloromethyl, pentafluoroethyl, pentachloroethyl,2,2,2-trifluoroethyl, heptafluoropropyl, and heptachloropropyl. Examplesof haloalkyl also include “fluoroalkyl” that is intended to include bothbranched and straight-chain saturated aliphatic hydrocarbon groupshaving the specified number of carbon atoms, substituted with 1 or morefluorine atoms. The term “polyhaloalkyl” as used herein refers to an“alkyl” group as defined above which includes from 2 to 9, preferablyfrom 2 to 5, halo substituents, such as F or C₁, preferably F, such aspolyfluoroalkyl, for example, CF₃CH₂, CF₃ or CF₃CF₂CH₂.

“Haloalkoxy” or “haloalkyloxy” represents a haloalkyl group as definedabove with the indicated number of carbon atoms attached through anoxygen bridge. For example, “C₁ to C₆ haloalkoxy” or “C₁₋₆ haloalkoxy”,is intended to include C₁, C₂, C₃, C₄, C₅, and C₆ haloalkoxy groups.Examples of haloalkoxy include, but are not limited to,trifluoromethoxy, 2,2,2-trifluoroethoxy, and pentafluorothoxy. Otherexamples of haloalkoxy also include “fluoroalkoxy” which represents afluoroalkyl group as defined above with the indicated number of carbonatoms attached through an oxygen bridge. Similarly, “haloalkylthio” or“thiohaloalkoxy” represents a haloalkyl group as defined above with theindicated number of carbon atoms attached through a sulphur bridge; forexample trifluoromethyl-S—, and pentafluoroethyl-S—. The term“polyhaloalkyloxy” as used herein refers to an “alkoxy” or “alkyloxy”group as defined above which includes from 2 to 9, preferably from 2 to5, halo substituents, such as F or C₁, preferably F, such aspolyfluoroalkoxy, for example, —OCH₂CF₃, —OCF₃, or —OCH₂CF₂CF₃.

“Hydroxyalkyl” is intended to include both branched and straight-chainsaturated aliphatic hydrocarbon groups having the specified number ofcarbon atoms, substituted with 1 or more hydroxyl (OH). “C₁ to C₆hydroxyalkyl” (or hydroxyalkyl), is intended to include C₁, C₂, C₃, C₄,C₅, and C₆ hydroxyalkyl groups.

The term “cycloalkyl” refers to cyclized alkyl groups, including mono-,bi- or poly-cyclic ring systems. “C₃ to C₇ cycloalkyl” or “C₃₋₇cycloalkyl” is intended to include C₃, C₄, C₅, C₆, and C₇ cycloalkylgroups. Example cycloalkyl groups include, but are not limited to,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and norbomyl. Branchedcycloalkyl groups such as 1-methylcyclopropyl and 2-methylcyclopropylare included in the definition of “cycloalkyl”.

The term “cycloheteroalkyl” refers to cyclized heteroalkyl groups,including mono-, bi- or poly-cyclic ring systems. “C₃ to C₇cycloheteroalkyl” or “C₃₋₇ cycloheteroalkyl” is intended to include C₃,C₄, C₅, C₆, and C₇ cycloheteroalkyl groups. Example cycloheteroalkylgroups include, but are not limited to, oxetanyl, tetrahydrofuranyl,tetrahydropyranyl, azetidinyl, pyrrolidinyl, piperidinyl, morpholinyl,and piperazinyl.

Branched cycloheteroalkyl groups, such as piperidinylmethyl,piperazinylmethyl, morpholinylmethyl, pyridinylmethyl, pyridizylmethyl,pyrimidylmethyl, and pyrazinylmethyl, are included in the definition of“cycloheteroalkyl”.

As used herein, the term “azacyclyl” refers to a cycloheteroalkylcontaining one or more nitrogen atoms in the ring. Example azacyclylgroups include, but are not limited to, pyrrolidinyl, piperidinyl,morpholinyl, and piperazinyl.

As used herein, “carbocycle”, “carbocyclyl”, or “carbocyclic” isintended to mean any stable 3-, 4-, 5-, 6-, 7-, or 8-membered monocyclicor 5-, 6-, 7-, 8-, 9-, 10-, 11-, 12-, or 13-membered polycyclic(including bicyclic or tricyclic) hydrocarbon ring, any of which may besaturated or partially unsaturated. That is, the term “carbocycle”,“carbocyclyl”, or “carbocyclic” includes, without limitation, cycloalkyland cycloalkenyl. Examples of such carbocycles include, but are notlimited to, cyclopropyl, cyclobutyl, cyclobutenyl, cyclopentyl,cyclopentenyl, cyclohexyl, cycloheptenyl, cycloheptyl, cycloheptenyl,adamantyl, cyclooctyl, cyclooctenyl, cyclooctadienyl,[3.3.0]bicyclooctane, [4.3.0]bicyclononane, [4.4.0]bicyclodecane(decalin), [2.2.2]bicyclooctane, fluorenyl, indanyl, adamantyl, andtetrahydronaphthyl (tetralin). As shown above, bridged rings are alsoincluded in the definition of carbocycle (e.g., [2.2.2]bicyclooctane).Preferred carbocycles, unless otherwise specified, are cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, indanyl, and tetrahydronaphthyl. Abridged ring occurs when one or more, preferably one to three, carbonatoms link two non-adjacent carbon atoms. Preferred bridges are one ortwo carbon atoms. It is noted that a bridge always converts a monocyclicring into a tricyclic ring. When a ring is bridged, the substituentsrecited for the ring may also be present on the bridge.

Furthermore, the term “carbocyclyl”, including “cycloalkyl” and“cycloalkenyl”, as employed herein alone or as part of another groupincludes saturated or partially unsaturated (containing 1 or 2 doublebonds) cyclic hydrocarbon groups containing 1 to 3 rings, includingmonocyclicalkyl, bicyclicalkyl and tricyclicalkyl, containing a total of3 to 20 carbons forming the rings, preferably 3 to 10 carbons or 3 to 6carbons, forming the ring and which may be fused to 1 or 2 aromaticrings as described for aryl, which include cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclodecyl andcyclododecyl, cyclohexenyl,

any of which groups may be optionally substituted with 1 to 4substituents such as halogen, alkyl, alkoxy, hydroxy, aryl, aryloxy,arylalkyl, cycloalkyl, alkylamido, alkanoylamino, oxo, acyl,arylcarbonylamino, nitro, cyano, thiol and/or alkylthio and/or any ofthe alkyl substituents.

As used herein, the term “bicyclic carbocycle” or “bicyclic carbocyclicgroup” is intended to mean a stable 9- or 10-membered carbocyclic ringsystem that contains two fused rings and consists of carbon atoms. Ofthe two fused rings, one ring is a benzo ring fused to a second ring;and the second ring is a 5- or 6-membered carbon ring which is saturatedor partially unsaturated. The bicyclic carbocyclic group may be attachedto its pendant group at any carbon atom which results in a stablestructure. The bicyclic carbocyclic group described herein may besubstituted on any carbon if the resulting compound is stable. Examplesof a bicyclic carbocyclic group are, but not limited to,1,2-dihydronaphthyl, 1,2,3,4-tetrahydronaphthyl, and indanyl.

As used herein, the term “aryl”, as employed herein alone or as part ofanother group, refers to monocyclic or polycyclic (including bicyclicand tricyclic) aromatic hydrocarbons, including, for example, phenyl,naphthyl, anthracenyl, and phenanthranyl. Aryl moieties are well knownand described, for example, in Lewis, R. J., ed., Hawley's CondensedChemical Dictionary, 13th Edition, John Wiley & Sons, Inc., New York(1997). In one embodiment, the term “aryl” denotes monocyclic andbicyclic aromatic groups containing 6 to 10 carbons in the ring portion(such as phenyl or naphthyl including 1-naphthyl and 2-naphthyl). Forexample, “C₆ or C₁₀ aryl” or “C₆₋₁₀ aryl” refers to phenyl and naphthyl.Unless otherwise specified, “aryl”, “C₆ or C₁₀ aryl”, “C₆₋₁₀ aryl”, or“aromatic residue” may be unsubstituted or substituted with 1 to 5groups, preferably 1 to 3 groups, selected from —OH, —OCH₃, F, C₁, Br,I, —CN, —NO₂, —NH₂, —N(CH₃)H, —N(CH₃)₂, —CF₃, —OCF₃, —C(O)CH₃, —SCH₃,—S(O)CH₃, —S(O)₂CH₃, —CH₃, —CH₂CH₃, —CO₂H, and —CO₂CH₃.

As used herein, the term “heterocycle”, “heterocyclyl”, or “heterocyclicgroup” is intended to mean a stable 3-, 4-, 5-, 6-, or 7-memberedmonocyclic or 5-, 6-, 7-, 8-, 9-, 10-, 11-, 12-, 13-, or 14-memberedpolycyclic (including bicyclic and tricyclic) heterocyclic ring that issaturated, or partially unsaturated, and that contains carbon atoms and1, 2, 3 or 4 heteroatoms independently selected from the groupconsisting of N, O and S; and including any polycyclic group in whichany of the above-defined heterocyclic rings is fused to a carbocyclic oran aryl (e.g., benzene) ring. That is, the term “heterocycle”,“heterocyclyl”, or “heterocyclic group” includes non-aromatic ringsystems, such as heterocycloalkyl and heterocycloalkenyl. The nitrogenand sulfur heteroatoms may optionally be oxidized (i.e., N→O andS(O)_(p), wherein p is 0, 1 or 2). The nitrogen atom may be substitutedor unsubstituted (i.e., N or NR wherein R is H or another substituent,if defined). The heterocyclic ring may be attached to its pendant groupat any heteroatom or carbon atom that results in a stable structure. Theheterocyclic rings described herein may be substituted on carbon or on anitrogen atom if the resulting compound is stable. A nitrogen in theheterocycle may optionally be quatemized. It is preferred that when thetotal number of S and O atoms in the heterocycle exceeds 1, then theseheteroatoms are not adjacent to one another. It is preferred that thetotal number of S and O atoms in the heterocycle is not more than 1.Examples of hetercyclyl include, without limitation, azetidinyl,piperazinyl, piperidinyl, piperidonyl, piperonyl, pyranyl, morpholinyl,tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl,morpholinyl, dihydrofuro[2,3-b]tetrahydrofuran.

As used herein, the term “bicyclic heterocycle” or “bicyclicheterocyclic group” is intended to mean a stable 9- or 10-memberedheterocyclic ring system which contains two fused rings and consists ofcarbon atoms and 1, 2, 3, or 4 heteroatoms independently selected fromthe group consisting of N, O and S. Of the two fused rings, one ring isa 5- or 6-membered monocyclic aromatic ring comprising a 5-memberedheteroaryl ring, a 6-membered heteroaryl ring or a benzo ring, eachfused to a second ring. The second ring is a 5- or 6-membered monocyclicring which is saturated, partially unsaturated, or unsaturated, andcomprises a 5-membered heterocycle, a 6-membered heterocycle or acarbocycle (provided the first ring is not benzo when the second ring isa carbocycle).

The bicyclic heterocyclic group may be attached to its pendant group atany heteroatom or carbon atom which results in a stable structure. Thebicyclic heterocyclic group described herein may be substituted oncarbon or on a nitrogen atom if the resulting compound is stable. It ispreferred that when the total number of S and O atoms in the heterocycleexceeds 1, then these heteroatoms are not adjacent to one another. It ispreferred that the total number of S and O atoms in the heterocycle isnot more than 1. Examples of a bicyclic heterocyclic group are, but notlimited to, 1,2,3,4-tetrahydroquinolinyl,1,2,3,4-tetrahydroisoquinolinyl, 5,6,7,8-tetrahydro-quinolinyl,2,3-dihydro-benzofuranyl, chromanyl, 1,2,3,4-tetrahydro-quinoxalinyl,and 1,2,3,4-tetrahydro-quinazolinyl.

Bridged rings are also included in the definition of heterocycle. Abridged ring occurs when one or more, preferably one to three, atoms(i.e., C, O, N, or S) link two non-adjacent carbon or nitrogen atoms.Examples of bridged rings include, but are not limited to, one carbonatom, two carbon atoms, one nitrogen atom, two nitrogen atoms, and acarbon-nitrogen group. It is noted that a bridge always converts amonocyclic ring into a tricyclic ring. When a ring is bridged, thesubstituents recited for the ring may also be present on the bridge.

As used herein, the term “heteroaryl” is intended to mean stablemonocyclic and polycyclic (including bicyclic and tricyclic) aromatichydrocarbons that include at least one heteroatom ring member such assulfur, oxygen, or nitrogen. Heteroaryl groups include, withoutlimitation, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl,furyl, quinolyl, isoquinolyl, thienyl, imidazolyl, thiazolyl, indolyl,pyrroyl, oxazolyl, benzofuryl, benzothienyl, benzthiazolyl, isoxazolyl,pyrazolyl, triazolyl, tetrazolyl, indazolyl, 1,2,4-thiadiazolyl,isothiazolyl, purinyl, carbazolyl, benzimidazolyl, indolinyl,benzodioxolanyl, and benzodioxane. Heteroaryl groups are substituted orunsubstituted. The nitrogen atom is substituted or unsubstituted (i.e.,N or NR wherein R is H or another substituent, if defined). The nitrogenand sulfur heteroatoms may optionally be oxidized (i.e., N→O andS(O)_(p), wherein p is 0, 1 or 2).

Examples of heteroaryl include, but are not limited to, acridinyl,azocinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl,benzothiophenyl, benzoxazolyl, benzoxazolinyl, benzthiazolyl,benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl,benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl,chromenyl, cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl,furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl,1H-indazolyl, imidazolopyridinyl, indolenyl, indolinyl, indolizinyl,indolyl, 3H-indolyl, isatinoyl, isobenzofuranyl, isochromanyl,isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl,isothiazolopyridinyl, isoxazolyl, isoxazolopyridinyl,methylenedioxyphenyl, naphthyridinyl, octahydroisoquinolinyl,oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl,1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolopyridinyl,oxazolidinylperimidinyl, oxindolyl, pyrimidinyl, phenanthridinyl,phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathianyl,phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrazinyl,pyrazolidinyl, pyrazolinyl, pyrazolopyridinyl, pyrazolyl, pyridazinyl,pyridooxazolyl, pyridoimidazolyl, pyridothiazolyl, pyridinyl,pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2-pyrrolidonyl, 2H-pyrrolyl,pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl,quinuclidinyl, tetrazolyl, tetrahydrofuranyl, tetrahydroisoquinolinyl,tetrahydroquinolinyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl,1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl,thianthrenyl, thiazolyl, thienyl, thiazolopyridinyl, thienothiazolyl,thienooxazolyl, thienoimidazolyl, thiophenyl, triazinyl,1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, andxanthenyl.

Examples of 5- to 10-membered heteroaryl include, but are not limitedto, pyridinyl, furanyl, thienyl, pyrazolyl, imidazolyl, imidazolidinyl,indolyl, tetrazolyl, isoxazolyl, oxazolyl, oxadiazolyl, oxazolidinyl,thiadiazinyl, thiadiazolyl, thiazolyl, triazinyl, triazolyl,benzimidazolyl, 1H-indazolyl, benzofuranyl, benzothiofuranyl,benztetrazolyl, benzotriazolyl, benzisoxazolyl, benzoxazolyl, oxindolyl,benzoxazolinyl, benzthiazolyl, benzisothiazolyl, isatinoyl,isoquinolinyl, octahydroisoquinolinyl, isoxazolopyridinyl, quinazolinyl,quinolinyl, isothiazolopyridinyl, thiazolopyridinyl, oxazolopyridinyl,imidazolopyridinyl, and pyrazolopyridinyl. Examples of 5- to 6-memberedheterocycles include, but are not limited to, pyridinyl, furanyl,thienyl, pyrrolyl, pyrazolyl, pyrazinyl, imidazolyl, imidazolidinyl,indolyl, tetrazolyl, isoxazolyl, oxazolyl, oxadiazolyl, oxazolidinyl,thiadiazinyl, thiadiazolyl, thiazolyl, triazinyl, and triazolyl.

Unless otherwise indicated, “carbocyclyl” or “heterocyclyl” includes oneto three additional rings fused to the carbocyclic ring or theheterocyclic ring (such as aryl, cycloalkyl, heteroaryl orcycloheteroalkyl rings, for example,

and may be optionally substituted through available carbon atoms with 1,2, or 3 groups selected from hydrogen, halo, haloalkyl, alkyl,haloalkyl, alkoxy, haloalkoxy, alkenyl, trifluoromethyl,trifluoromethoxy, alkynyl, cycloalkyl-alkyl, cycloheteroalkyl,cycloheteroalkylalkyl, aryl, heteroaryl, arylalkyl, aryloxy,aryloxyalkyl, arylalkoxy, alkoxycarbonyl, arylcarbonyl, arylalkenyl,aminocarbonylaryl, arylthio, arylsulfinyl, arylazo, heteroarylalkyl,heteroarylalkenyl, heteroarylheteroaryl, heteroaryloxy, hydroxy, nitro,cyano, thiol, alkylthio, arylthio, heteroarylthio, arylthioalkyl,alkoxyarylthio, alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl,arylaminocarbonyl, alkoxycarbonyl, aminocarbonyl, alkylcarbonyloxy,arylcarbonyloxy, alkylcarbonylamino, arylcarbonylamino, arylsulfinyl,arylsulfinylalkyl, arylsulfonylamino and arylsulfonaminocarbonyl and/orany of the alkyl substituents set out herein.

When any of the terms alkyl, alkenyl, alkynyl, cycloalkyl, carbocyclyl,heterocyclyl, aryl, and heteroaryl are used as part of another group,the number of carbon atoms and ring members are the same as thosedefined in the terms by themselves. For example, alkoxy, haloalkoxy,alkylamino, haloalkyl, hydroxyalkyl, aminoalkyl, haloalkoxy,alkoxyalkoxy, haloalkylamino, alkoxyalkylamino, haloalkoxyalkylamino,alkylthio, and the like each independently contains the number of carbonatoms which are the same as defined for the term “alkyl”, such as 1 to 4carbon atoms, 1 to 6 carbon atoms, 1 to 10 carbon atoms, etc.

Similarly, cycloalkoxy, heterocyclyloxy, cycloalkylamino,heterocyclylamino, aralkylamino, arylamino, aryloxy, aralkyloxy,heteroaryloxy, heteroarylalkyloxy, and the like each indepdentlycontains ring members which are the same as defined for the terms“cycloalkyl”, “heterocyclyl”, “aryl”, and “heteroaryl”, such as 3 to6-membered, 4 to 7-membered, 6 to 10-membered, 5 to 10-membered, 5 or6-membered, etc.

In accordance with a convention used in the art, a bond pointing to abold line, such as

as used in structural formulas herein, depicts the bond that is thepoint of attachment of the moiety or substituent to the core or backbonestructure.

In accordance with a convention used in the art, a wavy or squiggly bondin a structural formula, such as

is used to depict a stereogenic center of the carbon atom to which X′,Y′, and Z′ are attached and is intended to represent both enantiomers ina single figure. That is, a structural formula with such as wavy bonddenotes each of the enantiomers individually, such as

as well as a racemic mixture thereof. When a wavy or squiggly bond isattached to a double bond (such as C═C or C═N) moiety, it include cis-or trans- (or E- and Z-) geometric isomers or a mixture thereof.

It is understood herein that if a carbocyclic or heterocyclic moiety maybe bonded or otherwise attached to a designated substrate throughdiffering ring atoms without denoting a specific point of attachment,then all possible points are intended, whether through a carbon atom or,for example, a trivalent nitrogen atom. For example, the terms“pyridinyl” or “pyridyl” means 2-, 3- or 4-pyridinyl, the term “thienyl”means 2- or 3-thienyl, and so forth.

When a bond to a substituent is shown to cross a bond connecting twoatoms in a ring, then such substituent may be bonded to any atom on thering. When a substituent is listed without indicating the atom in whichsuch substituent is bonded to the rest of the compound of a givenformula, then such substituent may be bonded via any atom in suchsubstituent. Combinations of substituents and/or variables arepermissible only if such combinations result in stable compounds.

One skilled in the art will recognize that substituents and othermoieties of the compounds of the present invention should be selected inorder to provide a compound which is sufficiently stable to provide apharmaceutically useful compound which can be formulated into anacceptably stable pharmaceutical composition. Compounds of the presentinvention which have such stability are contemplated as falling withinthe scope of the present invention.

The term “counter ion” is used to represent a negatively charged speciessuch as chloride, bromide, hydroxide, acetate, and sulfate. The term“metal ion” refers to alkali metal ions such as sodium, potassium orlithium and alkaline earth metal ions such as magnesium and calcium, aswell as zinc and aluminum.

As referred to herein, the term “substituted” means that at least onehydrogen atom (attached to carbon atom or heteroatom) is replaced with anon-hydrogen group, provided that normal valencies are maintained andthat the substitution results in a stable compound. When a substituentis oxo (i.e., ═O), then 2 hydrogens on the atom are replaced. Oxosubstituents are not present on aromatic moieties. When a ring system(e.g., carbocyclic or heterocyclic) is said to be substituted with acarbonyl group or a double bond, it is intended that the carbonyl groupor double bond be part (i.e., within) of the ring. Ring double bonds, asused herein, are double bonds that are formed between two adjacent ringatoms (e.g., C═C, C═N, or N═N). The term “substituted” in reference toalkyl, cycloalkyl, heteroalkyl, cycloheteroalkyl, alkylene, aryl,arylalkyl, heteroaryl, heteroarylalkyl, carbocyclyl, and heterocyclyl,means alkyl, cycloalkyl, heteroalkyl, cycloheteroalkyl, alkylene, aryl,arylalkyl, heteroaryl, heteroarylalkyl, carbocyclyl, and heterocyclyl,respectively, in which one or more hydrogen atoms, which are attached toeither carbon or heteroatom, are each independently replaced with one ormore non-hydrogen substituent(s).

In cases wherein there are nitrogen atoms (e.g., amines) on compounds ofthe present invention, these may be converted to N-oxides by treatmentwith an oxidizing agent (e.g., mCPBA and/or hydrogen peroxides) toafford other compounds of this invention. Thus, shown and claimednitrogen atoms are considered to cover both the shown nitrogen and itsN-oxide (N→O) derivative.

When any variable occurs more than one time in any constituent orformula for a compound, its definition at each occurrence is independentof its definition at every other occurrence. Thus, for example, if agroup is shown to be substituted with 0, 1, 2, or 3 R groups, then saidgroup be unsubstituted when it is substituted with 0 R group, or besubstituted with up to three R groups, and at each occurrence R isselected independently from the definition of R.

Also, combinations of substituents and/or variables are permissible onlyif such combinations result in stable compounds.

As used herein, the term “tautomer” refers to each of two or moreisomers of a compound that exist together in equilibrium, and arereadily interchanged by migration of an atom or group within themolecule For example, one skilled in the art would readily understandthat a 1,2,3-triazole exists in two tautomeric forms as defined above:

Thus, this disclosure is intended to cover all possible tautomers evenwhen a structure depicts only one of them.

The phrase “pharmaceutically acceptable” is employed herein to refer tothose compounds, materials, compositions, and/or dosage forms that are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of human beings and animals without excessive toxicity,irritation, allergic response, and/or other problem or complication,commensurate with a reasonable benefit/risk ratio.

The compounds of the present invention can be present as salts, whichare also within the scope of this invention. Pharmaceutically acceptablesalts are preferred. As used herein, “pharmaceutically acceptable salts”refer to derivatives of the disclosed compounds wherein the parentcompound is modified by making acid or base salts thereof. Thepharmaceutically acceptable salts of the present invention can besynthesized from the parent compound that contains a basic or acidicmoiety by conventional chemical methods. Generally, such salts can beprepared by reacting the free acid or base forms of these compounds witha stoichiometric amount of the appropriate base or acid in water or inan organic solvent, or in a mixture of the two; generally, nonaqueousmedia like ether, ethyl acetate, ethanol, isopropanol, or acetonitrileare preferred. Lists of suitable salts are found in Remington'sPharmaceutical Sciences, 18th Edition, Mack Publishing Company, Easton,Pa. (1990), the disclosure of which is hereby incorporated by reference.

If the compounds of the present invention have, for example, at leastone basic center, they can form acid addition salts. These are formed,for example, with strong inorganic acids, such as mineral acids, forexample sulfuric acid, phosphoric acid or a hydrohalic acid, withorganic carboxylic acids, such as alkanecarboxylic acids of 1 to 4carbon atoms, for example acetic acid, which are unsubstituted orsubstituted, for example, by halogen as chloroacetic acid, such assaturated or unsaturated dicarboxylic acids, for example oxalic,malonic, succinic, maleic, fumaric, phthalic or terephthalic acid, suchas hydroxycarboxylic acids, for example ascorbic, glycolic, lactic,malic, tartaric or citric acid, such as amino acids, (for exampleaspartic or glutamic acid or lysine or arginine), or benzoic acid, orwith organic sulfonic acids, such as (C₁-C₄) alkyl or arylsulfonic acidswhich are unsubstituted or substituted, for example by halogen, forexample methyl- or p-toluene-sulfonic acid. Corresponding acid additionsalts can also be formed having, if desired, an additionally presentbasic center. The compounds of the present invention having at least oneacid group (for example COOH) can also form salts with bases. Suitablesalts with bases are, for example, metal salts, such as alkali metal oralkaline earth metal salts, for example sodium, potassium or magnesiumsalts, or salts with ammonia or an organic amine, such as morpholine,thiomorpholine, piperidine, pyrrolidine, a mono, di or tri-loweralkylamine, for example ethyl, tert-butyl, diethyl, diisopropyl,triethyl, tributyl or dimethyl-propylamine, or a mono, di or trihydroxylower alkylamine, for example mono, di or triethanolamine. Correspondinginternal salts may furthermore be formed. Salts which are unsuitable forpharmaceutical uses but which can be employed, for example, for theisolation or purification of free compounds of Formula (I) or theirpharmaceutically acceptable salts, are also included.

Preferred salts of the compounds of Formula (I) which contain a basicgroup include monohydrochloride, hydrogensulfate, methanesulfonate,phosphate, nitrate or acetate.

Preferred salts of the compounds of Formula (I) which contain an acidgroup include sodium, potassium and magnesium salts and pharmaceuticallyacceptable organic amines.

In addition, the compounds of the present invention may have prodrugforms. Any compound that will be converted in vivo to provide thebioactive agent is a prodrug within the scope and spirit of theinvention. The term “prodrug” as used herein encompasses both theprodrugs based on the carboxylic acid residue, i.e., “prodrug esters”,and the prodrugs based on the arginine mimetics moiety, i.e., “prodrugsof arginine mimetics”. Such prodrugs are preferably administered orallysince hydrolysis in many instances occurs principally under theinfluence of the digestive enzymes. Parenteral administration may beused where the esterper se is active, or in those instances wherehydrolysis occurs in the blood.

The compounds of the present invention contain a carboxy group which canform physiologically hydrolyzable esters that serve as prodrugs, i.e.,“prodrug esters”, by being hydrolyzed in the body to yield the compoundsof the present invention per se. Examples of physiologicallyhydrolyzable esters of compounds of the present invention include C₁ toC₆ alkyl, C₁ to C₆ alkylbenzyl, 4-methoxybenzyl, indanyl, phthalyl,methoxymethyl, C₁₋₆ alkanoyloxy-C₁₋₆ alkyl (e.g., acetoxymethyl,pivaloyloxymethyl or propionyloxymethyl), C₁ to C₆ alkoxycarbonyloxy-C₁to C₆ alkyl (e.g., methoxycarbonyl-oxymethyl or ethoxycarbonyloxymethyl,glycyloxymethyl, phenylglycyloxymethyl,(5-methyl-2-oxo-1,3-dioxolen-4-yl)-methyl), and other well knownphysiologically hydrolyzable esters used, for example, in the penicillinand cephalosporin arts. Such esters may be prepared by conventionaltechniques known in the art. The “prodrug esters” can be formed byreacting the carboxylic acid moiety of the compounds of the presentinvention with either alkyl or aryl alcohol, halide, or sulfonateemploying procedures known to those skilled in the art. Furthermore,various forms of prodrugs are well known in the art. For examples ofsuch prodrug derivatives, see:

-   Bundgaard, H., ed., Design of Prodrugs, Elsevier (1985), and    Widder, K. et al., eds., Methods in Enzymology, 112:309-396,    Academic Press (1985);-   Bundgaard, H., Chapter 5, “Design and Application of Prodrugs”,    Krosgaard-Larsen, P. et al., eds., A Textbook of Drug Design and    Development, pp. 113-191, Harwood Academic Publishers (1991);-   Bundgaard, H., Adv. Drug Deliv. Rev., 8:1-38 (1992);-   Bundgaard, H. et al., J. Pharm. Sci., 77:285 (1988); and-   Kakeya, N. et al., Chem. Pharm. Bull., 32:692 (1984).

Preparation of prodrugs is well known in the art and described in, forexample, King, F. D., ed., Medicinal Chemistry: Principles and Practice,The Royal Society of Chemistry, Cambridge, UK (1994); Testa, B. et al.,Hydrolysis in Drug and Prodrug Metabolism. Chemistry, Biochemistry andEnzymology, VCHA and Wiley-VCH, Zurich, Switzerland (2003); Wermuth, C.G., ed., The Practice of Medicinal Chemistry, Academic Press, San Diego,Calif. (1999); Rautio, J. et al., Nature Review Drug Discovery, 17,559-587, (2018).

The present invention is intended to include all isotopes of atomsoccurring in the present compounds. Isotopes include those atoms havingthe same atomic number but different mass numbers. By way of generalexample and without limitation, isotopes of hydrogen include deuterium(symbol D or ²H) and tritium (symbol T or ³H). Isotopes of carboninclude ¹³C and ¹⁴C. Isotopically-labeled compounds of the invention cangenerally be prepared by conventional techniques known to those skilledin the art or by processes analogous to those described herein, using anappropriate isotopically-labeled reagent in place of the non-labeledreagent otherwise employed. Such compounds have a variety of potentialuses, e.g., as standards and reagents in determining the ability of apotential pharmaceutical compound to bind to target proteins orreceptors, or for imaging compounds of this invention bound tobiological receptors in vivo or in vitro.

“Stable compound” and “stable structure” are meant to indicate acompound that is sufficiently robust to survive isolation to a usefuldegree of purity from a reaction mixture, and formulation into anefficacious therapeutic agent. It is preferred that compounds of thepresent invention do not contain a N-halo, S(O)₂H, or S(O)H group.

The term “solvate” means a physical association of a compound of thisinvention with one or more solvent molecules, whether organic orinorganic. This physical association includes hydrogen bonding. Thesolvent molecules in the solvate may be present in a regular arrangementand/or a non-ordered arrangement. The solvate may comprise either astoichiometric or nonstoichiometric amount of the solvent molecules.“Solvate” encompasses both solution-phase and isolable solvates.Exemplary solvates include, but are not limited to, hydrates,ethanolates, methanolates, and isopropanolates. Methods of solvation aregenerally known in the art.

The term “glycosyl” means a monovalent free radical or substituentmoiety obtained by removing the hemiacetal hydroxyl group from thecyclic form of a monosaccharide and, by extension, of a loweroligosaccharide. In one embodiment, the glycosyl group has the followingstructure:

Abbreviations as used herein, are defined as follows: “1×” for once,“2×” for twice, “3×” for thrice, “° C.” for degrees Celsius, “eq” forequivalent or equivalents, “g” for gram or grams, “mg” for milligram ormilligrams, “L” for liter or liters, “mL” for milliliter or milliliters,“μL” for microliter or microliters, “N” for normal, “M” for molar,“mmol” for millimole or millimoles, “min” for minute or minutes, “h” forhour or hours, “rt” for room temperature, “RBF” for round bottom flask,“atm” for atmosphere, “psi” for pounds per square inch, “conc.” forconcentrated, “RCM” for ring-closing metathesis, “sat” or “sat'd” forsaturated, “SFC” for supercritical fluid chromatography, “MW” formolecular weight, “mp” for melting point, “ee” for enantiomeric excess,“MS” or “Mass Spec” for mass spectrometry, “ESI” for electrosprayionization mass spectroscopy, “HR” for high resolution, “HRMS” for highresolution mass spectrometry, “LCMS” for liquid chromatography massspectrometry, “HPLC” for high pressure liquid chromatography, “RP HPLC”for reverse phase HPLC, “TLC” or “tlc” for thin layer chromatography,“NMR” for nuclear magnetic resonance spectroscopy, “nOe” for nuclearOverhauser effect spectroscopy, “¹H” for proton, “δ” for delta, “s” forsinglet, “d” for doublet, “t” for triplet, “q” for quartet, “m” formultiplet, “br” for broad, “Hz” for hertz, and “α”, “β”, “R”, “S”, “E”,and “Z” are stereochemical designations familiar to one skilled in theart.

Abbreviations

Abbreviations as used herein, are defined as follows: “1×” for once,“2×” for twice, “3×” for thrice, “° C.” for degrees Celsius, “eq” forequivalent or equivalents, “g” for gram or grams, “mg” for milligram ormilligrams, “L” for liter or liters, “mL” for milliliter or milliliters,“μL” for microliter or microliters, “N” for normal, “M” for molar,“mmol” for millimole or millimoles, “min” for minute or minutes, “h” forhour or hours, “rt” for room temperature, “RBF” for round bottom flask,“atm” for atmosphere, “psi” for pounds per square inch, “conc.” forconcentrated, “RCM” for ring-closing metathesis, “sat” or “sat'd” forsaturated, “SFC” for supercritical fluid chromatography, “MW” formolecular weight, “mp” for melting point, “ee” for enantiomeric excess,“MS” or “Mass Spec” for mass spectrometry, “ESI” for electrosprayionization mass spectroscopy, “HR” for high resolution, “HRMS” for highresolution mass spectrometry, “LCMS” for liquid chromatography massspectrometry, “HPLC” for high pressure liquid chromatography, “RP HPLC”for reverse phase HPLC, “TLC” or “tlc” for thin layer chromatography,“NMR” for nuclear magnetic resonance spectroscopy, “nOe” for nuclearOverhauser effect spectroscopy, “¹H” for proton, “δ” for delta, “s” forsinglet, “d” for doublet, “t” for triplet, “q” for quartet, “m” formultiplet, “br” for broad, “Hz” for hertz, and “α”, “β”, “R”, “S”, “E”,and “Z” are stereochemical designations familiar to one skilled in theart.

Furthermore, the following abbreviations are employed in the Schemes,Examples and elsewhere herein:

-   Me methyl-   Et Ethyl-   Pr propyl-   i-Pr isopropyl-   Bu Butyl-   i-Bu isobutyl-   t-Bu tert-butyl-   Ph phenyl-   Bn benzyl-   Boc or BOC tert-butyloxycarbonyl-   Boc₂O di-tert-butyl dicarbonate-   ACN acetonitrile-   AcOH or HOAc acetic acid-   AlCl₃ aluminum chloride-   AIBN Azobisisobutyronitrile-   BBr₃ boron tribromide-   BCl₃ boron trichloride-   BEMP    2-tert-butylimino-2-diethylamino-1,3-dimethylperhydro-1,3,2-diazaphosphorine-   BOP reagent benzotriazol-1-yloxytris(dimethylamino)phosphonium    hexafluorophosphate-   Burgess reagent 1-methoxy-N-triethylammoniosulfonyl-methanimidate-   CBz carbobenzyloxy-   DCM or CH₂Cl₂ dichloromethane-   CH₃CN or ACN acetonitrile-   CDCl₃ deutero-chloroform-   CHCl₃ chloroform-   mCPBA or m-CPBA meta-chloroperbenzoic acid-   Cs₂CO₃ cesium carbonate-   Cu(OAc)₂ copper (II) acetate-   Cy₂NMe N-cyclohexyl-N-methylcyclohexanamine-   DBU 1,8-diazabicyclo[5.4.0]undec-7-ene-   DCE 1,2 dichloroethane-   DEA diethylamine-   DMP or Dess-Martin Periodinane    1,1,1-tris(acetyloxy)-1,1-dihydro-1,2-beniziodoxol-3-(1H)-one-   DIC or DIPCDI diisopropylcarbodiimide-   DIEA, DIPEA or Hunig's base diisopropylethylamine-   DMAP 4-dimethylaminopyridine-   DME 1,2-dimethoxyethane-   DMF dimethyl formamide-   DMSO dimethyl sulfoxide-   cDNA complimentary DNA-   Dppp (R)-(+)-1,2-bis(diphenylphosphino)propane-   DuPhos (+)-1,2-bis((2S,5S)-2,5-diethylphospholano)benzene-   EDC N-(3-dimthylaminopropyl)-N′-ethylcarbodiimide-   EDCI N-(3-dimthylaminopropyl)-N′-ethylcarbodiimide hydrochloride-   EDTA ethylenediaminetetraacetic acid-   (S,S)-EtDuPhosRh(I)    (+)-1,2-bis((2S,5S)-2,5-diethylphospholano)benzene(1,5-cyclooctadiene)rhodium(I)    trifluoromethanesulfonate-   Et₃N or TEA triethylamine-   EtOAc ethyl acetate-   Et₂O diethyl ether-   EtOH ethanol-   GMF glass microfiber filter-   Grubbs II    (1,3-bis(2,4,6-trimethylphenyl)-2-imidazolidinylidene)dichloro(phenylmethylene)(triycyclohexylphosphine)ruthenium-   HCl hydrochloric acid-   HATU O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium    hexafluorophosphate-   HEPES 4-(2-hydroxyethyl)piperaxine-1-ethanesulfonic acid-   Hex hexanes-   HOBt or HOBT 1-hydroxybenzotriazole-   H₂O₂ hydrogen peroxide-   IBX 2-iodoxybenzoic acid-   H₂SO₄ sulfuric acid-   Jones reagent CrO₃ in aqueous H₂SO₄, 2 M-   K₂CO₃ potassium carbonate-   K₂HPO₄ potassium phosphate dibasic-   KOAc potassium acetate-   K₃PO₄ potassium phosphate-   LAH lithium aluminum hydride-   LG leaving group-   LiOH lithium hydroxide-   MeOH methanol-   MgSO₄ magnesium sulfate-   MsCl methanesulfonyl chloride-   MsOH or MSA methylsulfonic acid-   NaCl sodium chloride-   NaH sodium hydride-   NaHCO₃ sodium bicarbonate-   Na₂CO₃ sodium carbonate-   NaOH sodium hydroxide-   Na₂SO₃ sodium sulfite-   Na₂SO₄ sodium sulfate-   NBS N-bromosuccinimide-   NCS N-chlorosuccinimide-   NH₃ ammonia-   NH₄C₁ ammonium chloride-   NH₄OH ammonium hydroxide-   NH₄COOH ammonium formate-   NMM N-methylmorpholine-   OTf triflate or trifluoromethanesulfonate-   Pd₂(dba)₃ tris(dibenzylideneacetone)dipalladium(O)-   Pd(OAc)₂ palladium(II) acetate-   Pd/C palladium on carbon-   Pd(dppf)Cl₂    [1,1′-bis(diphenylphosphino)-ferrocene]dichloropalladium(II)-   Ph₃PCl₂ triphenylphosphine dichloride-   PG protecting group-   POCl₃ phosphorus oxychloride-   PPTS pyridinium p-toluenesulfonate-   i-PrOH or IPA isopropanol-   PS Polystyrene-   PtO₂ platinum oxide-   rt room temperature-   RuPhos-Pd-G2    chloro(2-dicyclohexylphosphino-2′,6′-diisopropoxy-1,1′-biphenyl)    [2-(2′-amino-1,1′-biphenyl)]palladium(II)-   SEM-Cl 2-(trimethysilyl)ethoxymethyl chloride-   SiO₂ silica oxide-   SnCl₂ tin(II) chloride-   TBAI tetra-n-butylammonium iodide-   TFA trifluoroacetic acid-   THF tetrahydrofuran-   TMSCHN₂ trimethylsilyldiazomethane-   T3P propane phosphonic acid anhydride-   TRIS tris(hydroxymethyl)aminomethane-   pTsOH p-toluenesulfonic acid-   TsCl p-tolunesulfonyl chloride

IV. Methods of Preparation

The compounds of the present invention can be prepared in a number ofways well known to one skilled in the art of organic synthesis using themethods described below, together with synthetic methods known in theart of synthetic organic chemistry, or variations thereon as appreciatedby those skilled in the art. Preferred methods include, but are notlimited to, those described below. All references cited herein arehereby incorporated in their entirety by reference. The reactions areperformed in a solvent or solvent mixture appropriate to the reagentsand materials employed and suitable for the transformations beingaffected. It will be understood by those skilled in the art of organicsynthesis that the functionality present on the molecule should beconsistent with the transformations proposed. This will sometimesrequire a judgment to modify the order of the synthetic steps or toselect one particular process scheme over another in order to obtain adesired compound of the invention. Restrictions to the substituents thatare compatible with the reaction conditions will be readily apparent toone skilled in the art and alternate methods must then be used. It willalso be recognized that another major consideration in the planning ofany synthetic route in this field is the judicious choice of theprotecting group used for protection of the reactive functional groupspresent in the compounds described in this invention. A particularlyuseful compendium of synthetic methods which may be applicable to thepreparation of compounds of the present invention may be found inLarock, R. C., Comprehensive Organic Transformations, VCH, New York(1989).

The compounds of the present invention may be prepared using thereactions and techniques described in this section. The reactions areperformed in solvents appropriate to the reagents and materials employedand are suitable for the transformations being effected. Also, in thedescription of the synthetic methods described below, it is to beunderstood that all proposed reaction conditions, including solvent,reaction atmosphere, reaction temperature, duration of the experimentand workup procedures, are chosen to be the conditions standard for thatreaction, which should be readily recognized by one skilled in the art.One skilled in the art of organic synthesis understands that thefunctionality present on various portions of the edict molecule must becompatible with the reagents and reactions proposed. Not all compoundsof Formula (I) falling into a given class may be compatible with some ofthe reaction conditions required in some of the methods described. Suchrestrictions to the substituents, which are compatible with the reactionconditions, will be readily apparent to one skilled in the art andalternate methods must be used. A particularly useful compendium ofsynthetic methods which may be applicable to the preparation ofcompounds of the present invention may be found in Larock, R. C.,Comprehensive Organic Transformations, VCH, New York (1989).

Generic Schemes

Compounds of the present invention, represented by Formula (I), Formula(II), Formula (III), or any subgenera or species thereof, can beprepared according to the general routes shown in SCHEMES 1 to 13 below.

Scheme 1 describes a method of preparing compounds of Formula I-a, I-b,and I-c, a subset of Formula I. Intermediate 1 can be converted toproducts I-a through coupling with X⁵-L¹-Z—R^(x) (X⁵ is a halogen,triflate or other suitable leaving group, and are commercially availableor readily prepared by methods known to one skilled in the art) underconditions that are well-known to one skilled in the art. In exampleswhere L¹ represents a covalent bond, products I-a can be obtainedthrough a variety of C—N bond forming reactions between intermediate 1and a suitable aryl halide, triflate or equivalent. Some examplesinclude, but are not limited to, Pd-catalyzed Buchwald-Hartwig reaction,Cu-mediated Ullmann coupling, Ni-mediated amination, or nucleophilicaromatic substitution (SNAr). Alternatively, the Cu-catalyzedChan-Evans-Lam coupling can be employed with a boronic acid or estercoupling partner. In each case, optimization of variables such ascatalyst, ligand, solvent, base, additives and temperature may berequired. In other examples L¹ represents a linker such as, but notlimited to, CO or SO₂. In such examples products I-a can be obtainedthrough the coupling of intermediate 1 with a suitable carboxylic acidutilizing coupling reagents such as but not limited to, T3P, EDC, DCC orCDI in the presence of a suitable base, for example triethylamine,Hunig's base, or pyridine with or without additives such as HOBT or DMAPin an appropriate solvent such as dichloromethane, ethyl acetate, DMF orTHF. In some examples, carboxylic acid chlorides or sulfonyl chloridesmay be reacted with intermediate 1 in order to obtain I-a by stirring inan appropriate solvent such as dichloromethane in the presence of a basesuch as triethylamine or Hunig's base. In each case the specificconditions utilized, including temperature, may require optimizationthat will be evident to one skilled in the art. If I-a contains an esteror nitrile it can be hydrolyzed to the corresponding carboxylic acid I-bunder conditions such as but not limited to treatment of I-a with NaOHor LiOH in solvents consisting of MeOH, THF, and water at a temperaturesuitable to enable the hydrolysis. Acid-mediated hydrolysis ofparticular esters, such as a tert-butyl ester, may be required in somecases to obtain I-b. Examples I-c can be obtained by the coupling of I-bwith R¹³—NH—R¹⁴ utilizing coupling reagents such as but not limited to,T3P, EDC, DCC or CDI in the presence of a suitable base, for exampletriethylamine, Hunig's base, or pyridine with or without additives suchas HOBT or DMAP in an appropriate solvent such as dichloromethane, ethylacetate, DMF or THF. In each case the specific conditions utilized toobtain I-c, including temperature and concentration, may requireoptimization.

Scheme 2 describes a method for the preparation of intermediates 1a, 1b,and 1c, a subset of intermediate 1. The coupling of intermediates 2,where X⁶ is Cl, Br or I, and ketones 3 can be accomplished through avariety of conditions such as formation of the aryl Grignard, aryllithium, aryl zinc or other aryl metal species of 2 with subsequentaddition to the ketone 3 to give tertiary alcohol products 4. Ketones 3are commercially available or can be prepared by methods that arewell-known to one skilled in the art. If appropriately acidic conditions(i.e. HCl, TFA) are employed during removal of the amino protectinggroup, for example where P*=Boc, alkenes 1a can be obtained as theprimary isolate (deprotection A). In other cases where P*=Boc, thehydroxyl can be retained to yield intermediates 1b if dilute or weaklyacidic conditions, such as TFA in DCM, are employed (deprotection B).Additionally, if P*=Cbz, palladium on carbon mediated hydrogenation canutilized to remove the protecting group without elimination of thehydroxyl to give 1b (deprotection B). Alkene intermediates 1a can bereduced under conditions such as, but not limited to, excesstriethylsilane heated in TFA as solvent to give intermediates 1c. Ifalternative protecting groups are required for functional groupcompatibility, then they can be removed by methods known to one skilledin the art. Additional methods for protecting group removal may be foundin Greene, T. and Wuts, P. G. M., Protecting Groups in OrganicSynthesis, John Wiley & Sons, Inc., New York, N.Y., 2006 and referencestherein.

Scheme 3 describes an alternative method for the synthesis ofintermediate 1a.

Ketones 3 can be converted to the corresponding boronic acid or ester intwo steps consisting of enol triflate formation and subsequent Miyauraborylation. Triflate formation can be accomplished by treating 3 with abase such as LiHMDS at low temperature in THF followed by addition ofComin's reagent or another suitable triflate donor source. Typicalconditions for Miyaura borylation include, but are not limited toheating the intermediate triflate with bis(pinacolato)diboron (B₂Pin₂),potassium acetate and a palladium catalyst such as PdCl₂(dppf)₂ in asuitable solvent such as THF or dioxane. Heteroaryl halide intermediate2 can undergo Suzuki coupling with boronic acid or boronic ester 5 toyield alkene 6. Typical conditions for the Suzuki coupling include, butare not limited to, heating the intermediates 2 and 5 together with apalladium catalyst, ligand and base at a suitable temperature in adeoxygenated solvent or solvent mixture. Specific conditions include,but are not limited to Pd(OAc)₂, DPEPhos, K₃PO₄ in dioxane/water at 90°C. In each case the specific conditions utilized to obtain 6, includingstoichiometry, palladium source, ligand, base, solvent, temperature, andconcentration may require independent optimization. Removal of theprotecting group P* can be accomplished as described in Scheme 2 to giveintermediate 1a.

Scheme 4 describes a method for preparing intermediate 1d, a subset ofintermediate 1. Intermediate 7 can undergo condensation with diol 8(commercially available or readily prepared by methods known to oneskilled in the art) under mildly-acidic dehydrating conditions to yieldacetal 9. Conditions for the conversion of 7 to 9 include, but are notlimited to, refluxing 7 and 8 in a solvent such as toluene in thepresence of 4 Å molecular sieves and catalyticp-TSA. Removal of theprotecting group P* in cases where P*=Boc, dilute or weakly acidicconditions, such as TFA in DCM, can be utilized to retain the acetal. Ifalternative protecting groups are required for functional groupcompatibility, then they can be removed by methods known to one skilledin the art. Additional methods for protecting group removal may be foundin Greene, T. and Wuts, P. G. M., Protecting Groups in OrganicSynthesis, John Wiley & Sons, Inc., New York, N.Y., 2006 and referencestherein.

Scheme 5 describes a method for preparing intermediate 1e, a subset ofintermediate 1. Intermediate aldehyde 7 can be converted to diol 10 intwo steps comprising alkenylation and dihydroxylation. The alkenylationstep can be accomplished with a reagent such as methyltriphenylphosphonium bromide and a suitable base such as, but notlimited to KOtBu or NaHMDS in a solvent such as THF. Subjecting theresultant alkene to conditions such as but not limited to OsO₄ and NMOin a suitable solvent gives diol 10. Condensation of 10 with ketone 11(commercially available or readily prepared by methods known to oneskilled in the art), under conditions such as catalytic p-TSA inrefluxing toluene with a drying agent such as 4 Å molecular sieves,yield ketal 12. Removal of the protecting group P* to give intermediate1e can be carried out as described in Scheme 4.

Alternatively, scheme 6 describes a method for the preparation ofcompounds I-d and I-e a subset of formula I. Intermediate 14 can beobtained through a variety of coupling reactions between amino ketone 13(commercially available or readily prepared by methods known to oneskilled in the art) and a suitable aryl halide, triflate or equivalentX⁵-L¹-Z—R^(x), where X⁵ represents the halide or triflate. Some examplesof such coupling reactions include, but are not limited to, Pd-catalyzedBuchwald-Hartwig reaction, Cu-mediated Ullmann coupling, Ni-mediatedamination, or nucleophilic aromatic substitution (SNAr) to giveintermediate 14. Subsequent condensation between ketone 14 and diol 10(scheme 5) under conditions that include mixing the reactants in thepresence of an acid catalyst like p-TSA in a solvent such as DCE, cangive products I-d. If products I-d contain an ester or nitrile they canbe hydrolyzed to the corresponding carboxylic acid I-e under conditionssuch as but not limited to treatment of I-d with NaOH or LiOH insolvents consisting of MeOH, THF, and water at a temperature suitable toenable the hydrolysis. Acid-mediated hydrolysis of particular esters,such as a tert-butyl ester, may be required in some cases to obtain I-e.

Scheme 7 describes a method for preparing intermediate 2a, a subset ofintermediate 2. Aldehydes 15 (commercially available or readily preparedby methods known to one skilled in the art) can be condensed withhydroxylamine hydrochloride under a variety of conditions including, butnot limited to, stirring both reactants in pyridine at room temperature,or gently heating the reactants in the presence of a base like sodiumhydroxide or sodium acetate in a suitable solvent such as ethanol. Theresultant oximes can be converted to the corresponding hydroximoylhalides 16 through halogenation by reagents such as but not limited toNCS or NBS in a suitable solvent such as DMF. The hydroximoyl halides 16undergo annulation with terminal alkynes (commercially available orreadily prepared by one skilled in the art) under conditions such as,but not limited to triethyl amine in dichloromethane at room temperatureto afford 3,5-substituted isoxazoles 17. The 4-position of the isoxazolecan be halogenated by reagents such as but not limited to NBS or NCS ina suitable solvent such as DMF to give 3,4,5-substituted isoxazoleintermediates 2a.

Scheme 8 describes a method of preparing intermediate 2b, a subset ofintermediate 2. The synthesis can commence with azidation of amine 18a(commercially available or readily prepared by methods known to oneskilled in the art) under conditions such as, but not limited to,treatment with sodium nitrite in acidic media (H₂O/TFA) followed byaddition of sodium azide in an appropriate solvent at a suitabletemperature to give azide 19. Alternatively, azide 19 can be obtained bythe reaction of halide 18b (commercially available or readily preparedby methods known to one skilled in the art) with an azide salt, such assodium azide, in a mixture of DMSO/water at an appropriate temperature.The resultant azide 19 can be annulated with a commercially availableterminal alkyne to give iodotriazole intermediate 2b under conditionssuch as, but not limited to, copper (II) perchlorate, potassium iodide,and DBU in THF at room temperature.

Scheme 9 describes a method of preparing intermediate 2c, a subset ofintermediate 2. Commercially available or readily prepared N-substitutedglycines 20 give sydnones 21 when treated with sodium nitrite, HCl andacetic anhydride under conditions that can be found in Fang, Y.; Wu, C.;Larock, R. C.; Shi, F. J. Org. Chem. 2011, 76, 8840. The sydnones 21 canbe converted to pyrazole intermediates 2c in a two-step processinvolving bromination with NBS followed by copper catalyzedcycloaddition with an alkyne as described in Decuypere, E.; Specklin,S.; Gabillet, S.; Audisio, D.; Liu, H.; Plougastel, L.; Kolodych S.;Taran, F. Org. Lett. 2015, 17, 362.

Scheme 10 describes a method for preparing intermediate 4a, a subset ofintermediate 4. An appropriately substituted boronic acid or ester 22(commercially available or readily prepared by methods known to oneskilled in the art) and a pyrazole 23 bearing a suitably reactivehalogen or equivalent X, (commercially available or readily prepared bymethods known to one skilled in the art) can be coupled through thePd-catalyzed Suzuki reaction to give intermediate 24. Typical conditionsfor the Suzuki coupling include, but are not limited to, heating thereactants 22 and 23 together with a palladium catalyst, ligand and baseat a suitable temperature in a deoxygenated solvent or solvent mixture.Specific conditions include, but are not limited to PdCl₂(dppf)₂, Na₂CO₃in THF/water at 120° C. In each case the specific conditions utilized toobtain 24, including stoichiometry, palladium source, ligand, base,solvent, temperature, and concentration may require independentoptimization. The coupling partners 22 and 23, are either commerciallyavailable or can be readily prepared by methods known to one skilled inthe art. Intermediate 24 can be deprotonated at the 5-position of thepyrazole by a sufficiently strong base such as, but not limited to,n-BuLi, or LDA in a suitable solvent such as THF or Et₂O. The resultinganion from deprotonation of 24 can be trapped in situ with a ketone 3 togive intermediate 2a.

Scheme 11 describes a method of preparing intermediate 7a, a subset ofintermediate 7. Hydroximoyl halides 16 (preparation described in Scheme6) can be reacted with β-ketoesters (commercially available or readilyprepared by methods known to one skilled in the art) in the presence oftriethyl amine or another suitable base in a solvent such as, but notlimited to, DCM to give 3,4,5-substituted isoxazole esters 25. Reductionof the ester can be accomplished by a number of reagents including, butnot limited to LiAlH₄, DIBAL-H, or LiBH₄ in an appropriate solvent. Theresultant hydroxyl of isoxazole 26 can be converted to aldehydeintermediates 7a under oxidative conditions including, but not limitedto PCC oxidation, Dess-Martin oxidation, Swem oxidation, Ley oxidationin an appropriate solvent such as, but not limited to DCM or DCE.

Scheme 12 describes a method of preparing compounds I-f, I-g, I-h asubset of Formula I. In some examples L¹ represents a linker atom suchas, but not limited to, O, or N and products I-f can be obtained throughthe coupling of intermediate 27 with X⁵—Z—R^(x) (X⁵ represents a halideor triflate) under conditions that include, but are not limited to,nucleophilic aromatic substitution (SNAr), transition metal mediatedarylation (i.e. Pd, Cu, Ni), Mitsunobu coupling, reductive amination oralkylation. If I-f contains an ester or nitrile it can be hydrolyzed tothe corresponding carboxylic acid I-g under conditions such as but notlimited to treatment with NaOH or LiOH in solvents consisting of MeOH,THF, and water at a temperature suitable to enable the hydrolysis.Acid-mediated hydrolysis of particular esters, such as a tert-butylester, may be required in some cases to obtain I-g. Examples I-h can beobtained by the coupling of I-g with R¹³—NH—R¹⁴ utilizing couplingreagents such as but not limited to, T3P, EDC, DCC or CDI in thepresence of a suitable base, for example triethylamine, Hunig's base, orpyridine with or without additives such as HOBT or DMAP in anappropriate solvent such as dichloromethane, ethyl acetate, DMF or THF.In each case the specific conditions utilized to obtain I-f, I-g, andI-h including temperature and concentration, may require optimization.

Scheme 13 describes a method of preparing compounds I-i, I-j, I-k asubset of Formula I. Products I-i can be obtained through coupling ofintermediate 28 with X⁵—Z—R^(x) (X⁵ represents a halide or triflate)under conditions that include, formation of the aryl Grignard, aryllithium, aryl zinc or other aryl metal species of X⁵—Z—R^(x) withsubsequent addition to 28. If I-i contains an ester or nitrile it can behydrolyzed to the corresponding carboxylic acid I-j under conditionssuch as but not limited to treatment with NaOH or LiOH in solventsconsisting of MeOH, THF, and water at a temperature suitable to enablethe hydrolysis. Acid-mediated hydrolysis of particular esters, such as atert-butyl ester, may be required in some cases to obtain I-j. ExamplesI-k can be obtained by the coupling of I-j with R¹³—NH—R¹⁵ utilizingcoupling reagents such as but not limited to, T3P, EDC, DCC or CDI inthe presence of a suitable base, for example triethylamine, Hunig'sbase, or pyridine with or without additives such as HOBT or DMAP in anappropriate solvent such as dichloromethane, ethyl acetate, DMF or THF.In each case the specific conditions utilized to obtain I-i, I-j, andI-k including temperature and concentration, may require optimization.

Scheme 14 describes a method for the preparation of intermediates 27a,27b, and 27c, a subset of intermediate 27. The coupling of heteroarylhalide intermediate 2 and ketone 29 (commercially available or readilyprepared by methods known to one skilled in the art) can be accomplishedthrough a variety of conditions such as formation of the aryl Grignard,aryl lithium, aryl zinc or other aryl metal species of 2 with subsequentaddition to the ketone 29 to give tertiary alcohol products 30. Ifappropriately acidic conditions (i.e. HCl, TFA) are employed duringremoval of the amino protecting group, for example where P*=Boc, alkenes27a can be obtained as the primary isolate (deprotection A). In othercases where P*=Boc, the hydroxyl can be retained to yield intermediates27b if dilute or weakly acidic conditions, such as TFA in DCM, areemployed. Additionally, if P*=Cbz, palladium on carbon mediatedhydrogenation can utilized to remove the protecting group withoutelimination of the hydroxyl to give 27b. Alkene intermediates 27a can bereduced under conditions such as, but not limited to, excesstriethylsilane heated in TFA as solvent to give intermediates 27c. Ifalternative protecting groups are required for functional groupcompatibility, then they can be removed by methods known to one skilledin the art. Additional methods for protecting group removal may be foundin Greene, T. and Wuts, P. G. M., Protecting Groups in OrganicSynthesis, John Wiley & Sons, Inc., New York, N.Y., 2006 and referencestherein.

EXAMPLES

The following Examples are offered as illustrative, as a partial scopeand particular embodiments of the invention and are not meant to belimiting of the scope of the invention. Abbreviations and chemicalsymbols have their usual and customary meanings unless otherwiseindicated. Unless otherwise indicated, the compounds described hereinhave been prepared, isolated and characterized using the schemes andother methods disclosed herein or may be prepared using the same.

As appropriate, reactions were conducted under an atmosphere of drynitrogen (or argon). For anhydrous reactions, DRISOLV® solvents from EMwere employed. For other reactions, reagent grade or HPLC grade solventswere utilized. Unless otherwise stated, all commercially obtainedreagents were used as received.

HPLC/MS and Preparatory/Analytical HPLC Methods Employed inCharacterization or Purification of Examples

NMR (nuclear magnetic resonance) spectra were typically obtained onBruker or JEOL 400 MHz and 500 MHz instruments in the indicatedsolvents. All chemical shifts are reported in ppm from tetramethylsilanewith the solvent resonance as the internal standard. ¹HNMR spectral dataare typically reported as follows: chemical shift, multiplicity(s=singlet, br s=broad singlet, d=doublet, dd=doublet of doublets,t=triplet, q=quartet, sep=septet, m=multiplet, app=apparent), couplingconstants (Hz), and integration.

The term HPLC refers to a Shimadzu high performance liquidchromatography instrument with one of following methods:

General Method A Example 12-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

Step 1. 2,6-Dichlorobenzaldehyde oxime

Hydroxylamine hydrochloride (6.6 g, 95 mmol) was added to a roomtemperature solution of 2,6-dichlorobenzaldehyde (11.1 g, 63.4 mmol) inpyridine (31.7 mL) giving a mild exotherm. After 10 minutes the excesspyridine was removed in vacuo and the residue was partitioned betweenEt₂O and water. The organic layer was sequentially washed with saturatedaqueous NH₄Cl, brine and the combined aqueous layers were back extractedwith several small portions of Et₂O. The combined organic extracts weredried over Na₂SO₄, filtered and concentrated in vacuo to give a2,6-dichlorobenzaldehyde oxime (12.4 g, 65.3 mmol, 100% yield) as awhite solid. The product was carried on to the next step without furtherpurification. ¹H NMR (400 MHz, CDCl₃) δ 8.39 (s, 1H), 7.92 (s, 1H),7.40-7.36 (m, 2H), 7.27-7.22 (m, 1H).

Step 2. 2,6-Dichloro-N-hydroxybenzimidoyl chloride

2,6-Dichlorobenzaldehyde oxime (12.0 g, 63.1 mmol) was dissolved in DMF(45.9 mL) and heated to 40° C. NCS (10.1 g, 76.0 mmol) dissolved in DMF(38.3 mL) was then added to the warm solution over the space ofapproximately 3 minutes. After stirring overnight at 40° C. the reactionmixture was cooled to room temperature, poured into ice, and extractedwith Et₂O. The organic layer was collected and washed with brine. Thecombined aqueous layers were back extracted with Et₂O. The combinedorganic layers were dried over Na₂SO₄, filtered and concentrated todryness in vacuo. The residue was purified by flash chromatography onSiO₂ (0-50% EtOAc/hexanes, Isco 120 g column) to give2,6-dichloro-N-hydroxybenzimidoyl chloride (13.3 g, 59.3 mmol, 94%yield) as a waxy white solid. ¹H NMR (500 MHz, CDCl₃) δ 8.02 (s, 1H),7.43-7.37 (m, 2H), 7.37-7.30 (m, 1H).

Step 3. 5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazole

Cyclopropylacetylene (2.8 mL, 33.4 mmol) followed by Et₃N (3.7 mL, 26.7mmol) were added to a room temperature solution of2,6-dichloro-N-hydroxybenzimidoyl chloride (5.0 g, 22.3 mmol) in DCM(111 mL). The reaction mixture was stirred at room temperature overnightand was concentrated onto SiO₂ for purification. The resulting mixturewas purified by flash chromatography on SiO₂ (0-45% EtOAc/hexanes, Isco120 g column) to give 5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazole (4.8g, 18.9 mmol, 85% yield) as a white solid. ¹H NMR (400 MHz, CDCl₃) δ7.43-7.39 (m, 2H), 7.34-7.28 (m, 1H), 6.01 (s, 1H), 2.13 (tt, J=8.2, 5.3Hz, 1H), 1.16-1.07 (m, 4H).

Step 4. 4-Bromo-5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazole

N-Bromosuccinimide (0.81 g, 4.6 mmol) was added to a room temperaturesolution of 5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazole (0.93 g, 3.7mmol) in DMF (14.6 mL). The reaction mixture was heated to 50° C. Afterheating overnight, additional N-bromosuccinimide (0.81 g, 4.6 mmol) wasadded and heating was continued. After heating for an additional 24hours the reaction was cooled to room temperature and poured intoapproximately 100 mL of ice water. The resulting solid was collected bysuction filtration and dried under high vacuum to give4-bromo-5-cyclopropyl-3-(2,6-dichlorophenyl) isoxazole (1.14 g, 3.42mmol, 94% yield) as a white powder. ¹H NMR (400 MHz, CDCl₃) δ 7.49-7.36(m, 3H), 2.19 (tt, J=8.4, 5.1 Hz, 1H), 1.36-1.29 (m, 2H), 1.24-1.16 (m,2H).

Step 5. tert-Butyl2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-2-hydroxy-7-azaspiro[3.5]nonane-7-carboxylate

n-Butyllithium (8.1 mL, 20.3 mmol) was added slowly to a −78° C.solution of 4-bromo-5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazole (5.4g, 16.2 mmol) in THF (64.9 mL) giving a light brown solution. After 10minutes, tert-butyl 2-oxo-7-azaspiro[3.5]nonane-7-carboxylate (3.9 g,16.2 mmol) was added as a solution in 3 mL of THF. The temperature wasmaintained at −78° C. for 3 hours. The cold reaction mixture wasquenched by the slow addition of 5 mL of methanol and then concentratedonto SiO₂ for purification by flash chromatography on SiO₂ (0-80%EtOAc/hexanes, Isco 120 g column) to give tert-butyl2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-2-hydroxy-7-azaspiro[3.5]nonane-7-carboxylate(5.4 g, 10.9 mmol, 67% yield) as a white powder. ¹H NMR (500 MHz, CDCl₃)δ 7.47-7.35 (m, 3H), 3.32-3.25 (m, 2H), 3.23-3.16 (m, 2H), 2.30 (s, 1H),2.07-2.00 (m, 2H), 1.70 (br d, J=1.4 Hz, 2H), 1.46 (br t, J=3.0 Hz, 2H),1.43 (s, 8H), 1.41-1.35 (m, 2H), 1.32-1.24 (m, 3H), 1.18-1.12 (m, 2H).

Step 6.5-Cyclopropyl-3-(2,6-dichlorophenyl)-4-(7-azaspiro[3.5]non-1-en-2-yl)isoxazole

Trifluoroacetic acid (8.6 mL, 111.0 mmol) was added to a flaskcontaining tert-butyl2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-2-hydroxy-7-azaspiro[3.5]nonane-7-carboxylate(5.5 g, 11.2 mmol). The mixture was stirred at room temperature for onehour and the excess TFA was removed in vacuo. The residue was dilutedwith EtOAc and washed with saturated aqueous K₂CO₃ and then brine. Thecombined aqueous layers were back extracted with EtOAc and the combinedorganic extracts were dried over Na₂SO₄, filtered and concentrated invacuo to dryness giving5-cyclopropyl-3-(2,6-dichlorophenyl)-4-(7-azaspiro[3.5]non-1-en-2-yl)isoxazole(4.2 g, 11.2 mmol, 100% yield) as a white solid. ¹H NMR (400 MHz,DMSO-d₆) δ 8.43-8.24 (m, 1H), 7.71-7.57 (m, 3H), 5.89 (s, 1H), 3.33 (brs, 2H), 3.06 (br s, 2H), 3.00-2.88 (m, 2H), 2.35 (s, 1H), 2.34-2.25 (m,1H), 1.72-1.63 (m, 3H), 1.27-1.11 (m, 4H).

Example 1.2-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

Cesium carbonate (0.1 g, 0.33 mmol) and ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate (60.8 mg, 0.20 mmol) wereadded to a room temperature solution of5-cyclopropyl-3-(2,6-dichlorophenyl)-4-(7-azaspiro[3.5]non-1-en-2-yl)isoxazole(50 mg, 0.13 mmol) in DMA (0.38 mL), and the reaction mixture was heatedto 90° C. After heating for 2 hours the reaction mixture was dilutedwith THF (1.0 mL), water (0.2 mL) and MeOH (0.1 mL). Lithium hydroxidemonohydrate (0.02 g, 0.40 mmol) was added to the mixture and thereaction vessel was sealed and heated to 90° C. overnight. The reactionwas then quenched by the addition of 0.5 mL of 1.0 N HCl. The resultantmixture was loaded onto a pad of Celite in an Isco dry load cartridgefor purification by C-18 reverse phase flash chromatography (10-100% Bin A, A=10:90:0.1 MeCN:H₂O:TFA, B=90:10:0.1 MeCN:H₂O:TFA, 18 min lineargradient, Isco 50 g C-18 gold column) desired fractions were combinedand concentrated to give2-(2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid (69 mg, 0.1 mmol, 88% yield) as a tan solid. MS (ESI) m/z: 570.6[M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δ 8.12 (d, J=1.5 Hz, 1H), 7.74 (dd,J=11.1, 1.4 Hz, 1H), 7.47-7.40 (m, 2H), 7.39-7.33 (m, 1H), 5.78 (s, 1H),3.78 (dt, J=13.1, 5.1 Hz, 2H), 3.63-3.51 (m, 2H), 2.43 (s, 2H), 2.18(tt, J=8.4, 5.0 Hz, 1H), 1.80-1.74 (m, 4H), 1.38-1.28 (m, 2H), 1.21-1.11(m, 2H); FXR EC₅₀=7 nM; Mouse in vivo (3 mg/kg, @ 6 h): Cyp7a1=−99%,Fgf15 =+18×; (30 mg/kg, @ 6 h): Cyp7a1=−99%, Fgf15=+31×.

Example 22-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl6-fluoronicotinate. MS (ESI) m/z: 496.1 [M+H]⁺; ¹H NMR (500 MHz,DMSO-d₆) δ 8.57 (d, J=1.8 Hz, 1H), 7.87 (dd, J=9.0, 2.3 Hz, 1H),7.68-7.61 (m, 2H), 7.61-7.54 (m, 1H), 6.81 (d, J=9.2 Hz, 1H), 5.85 (s,1H), 3.86-3.75 (m, 2H), 3.42-3.30 (m, 1H), 2.38-2.25 (m, 3H), 1.58-1.46(m, 4H), 1.27-1.16 (m, 2H), 1.15-1.07 (m, 2H); FXR EC₅₀=31 nM.

Example 32-(3-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-1-oxa-8-azaspiro[4.5]dec-3-en-8-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of tert-butyl2-oxo-7-azaspiro[3.5]nonane-7-carboxylate with tert-butyl3-oxo-1-oxa-8-azaspiro[4.5]decane-8-carboxylate. MS (ESI) m/z: 586.3[M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ 8.14 (s, 1H), 7.70-7.51 (m, 4H),5.67 (s, 1H), 4.57 (s, 2H), 3.74 (br s, 1H), 3.50 (br t, J=10.8 Hz, 2H),2.34-2.23 (m, 1H), 1.78-1.66 (m, 2H), 1.59 (br d, J=13.4 Hz, 2H),1.25-1.16 (m, 2H), 1.13-1.06 (m, 2H); FXR EC₅₀=240 nM.

Example 46-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)picolinic acid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl6-fluoropicolinate. MS (ESI) m/z: 495.8 [M+H]⁺; ¹H NMR (500 MHz,DMSO-d₆) δ 7.70-7.57 (m, 4H), 7.24 (d, J=7.0 Hz, 1H), 7.02 (d, J=8.9 Hz,1H), 5.85 (s, 1H), 3.85-3.69 (m, 2H), 3.41-3.23 (m, 1H), 2.38-2.27 (m,3H), 1.54 (br s, 4H), 1.28-1.17 (m, 3H), 1.17-1.09 (m, 3H); FXR EC₅₀=712nM.

Example 56-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)pyridazine-3-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl6-chloropyridazine-3-carboxylate. MS (ESI) m/z: 497.2 [M+H]⁺; ¹H NMR(500 MHz, DMSO-d₆) δ 7.76 (br d, J=9.5 Hz, 1H), 7.70-7.61 (m, 2H),7.61-7.53 (m, 1H), 7.21 (br d, J=9.8 Hz, 1H), 5.86 (s, 1H), 3.88 (br d,J=13.1 Hz, 1H), 3.44 (br d, J=4.6 Hz, 1H), 2.34 (s, 3H), 1.91 (s, 1H),1.57 (br s, 4H), 1.29-1.16 (m, 2H), 1.13 (br d, J=2.1 Hz, 2H) FXREC₅₀=313 nM.

Example 62-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-7-methyl-7H-pyrrolo[2,3-d]pyrimidine-5-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl2-chloro-7-methyl-7H-pyrrolo[2,3-d]pyrimidine-5-carboxylate. MS (ESI)m/z: 550.1 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ 8.78 (s, 1H), 7.75 (s,1H), 7.67-7.60 (m, 2H), 7.61-7.53 (m, 1H), 5.85 (s, 1H), 3.99 (br d,J=13.4 Hz, 1H), 3.81-3.67 (m, 2H), 3.53-3.39 (m, 1H), 2.40-2.22 (m, 3H),1.89 (s, 1H), 1.52 (br s, 4H), 1.29-1.16 (m, 2H), 1.11 (br d, J=2.4 Hz,2H) additional peaks under DMSO and H₂O peaks; FXR EC₅₀=47 nM.

General Method B Example 76-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-1-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylicacid

A slurry of5-cyclopropyl-3-(2,6-dichlorophenyl)-4-(7-azaspiro[3.5]non-1-en-2-yl)isoxazole(0.13 g, 0.34 mmol, synthesis described in General Method A), methyl6-chloro-1-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylate (77 mg, 0.34mmol) and Cs₂CO₃ (0.22 g, 0.69 mmol) in dioxane (3.4 mL) was degassed bybubbling nitrogen through the mixture for 5 minutes.Chloro(2-dicyclohexylphosphino-2′,6′-diisopropoxy-1,1′-biphenyl)[2-(2′-amino-1,1′-biphenyl)]palladium(II)(RuPhos-Pd-G2) (13.3 mg, 0.02 mmol) was then added and the reactionmixture was sealed and heated to 90° C. After heating overnightadditional RuPhos-Pd-G2 (13.3 mg, 0.02 mmol) was added, nitrogen wasbubbled through the mixture and it was resealed and heated to 100° C.After 1 hour the reaction mixture was concentrated to dryness and theresidue was dissolved in a mixture of THF (1.0 mL), water (0.4 mL), andMeOH (0.1 mL). Lithium hydroxide monohydrate (27.9 mg, 0.67 mmol) wasadded to the mixture and the reaction vessel was sealed and heated to90° C. After heating for 2 hours the reaction was quenched with 1N HCland then concentrated in vacuo to minimal volume. The residue was takenup in MeOH, filtered, and the crude material was purified viapreparative LC/MS with the following conditions: Column: XBridge C18,19×200 mm, 5-μm particles; Mobile Phase A: 5:95 acetonitrile: water with10-mM ammonium acetate; Mobile Phase B: 95:5 acetonitrile: water with10-mM ammonium acetate; Gradient: 45-100% B over 24 minutes, then a10-minute hold at 100% B; Flow: 20 mL/min. Fractions containing thedesired product were combined and dried via centrifugal evaporation togive6-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-1-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylicacid. MS (ESI) m/z: 549.2 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 8.00 (d,J=8.8 Hz, 1H), 7.80 (s, 1H), 7.67 (d, J=1.8 Hz, 1H), 7.65 (d, J=0.7 Hz,1H), 7.62-7.56 (m, 1H), 6.79 (d, J=8.8 Hz, 1H), 5.84 (s, 1H), 3.81-3.71(m, 3H), 3.31 (td, J=8.5, 3.5 Hz, 2H), 2.38-2.27 (m, 5H), 1.66-1.51 (m,5H), 1.25-1.17 (m, 2H), 1.16-1.07 (m, 3H); FXR EC₅₀=24 nM.

Example 86-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-1-methyl-1H-indole-3-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 7 with replacement of methyl6-chloro-1-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylate with methyl6-bromo-1-methyl-1H-indole-3-carboxylate. MS (ESI) m/z: 548.2 [M+H]⁺; ¹HNMR (500 MHz, DMSO-d₆) δ 7.86-7.75 (m, 2H), 7.71-7.63 (m, 2H), 7.63-7.55(m, 1H), 6.97-6.86 (m, 2H), 5.87 (s, 1H), 3.75 (s, 3H), 3.30-3.20 (m,1H), 2.96 (br t, J=9.2 Hz, 2H), 2.40-2.28 (m, 3H), 1.77-1.68 (m, 2H),1.68-1.57 (m, 2H), 1.28-1.16 (m, 3H), 1.14 (br d, J=2.7 Hz, 2H); FXREC₅₀=45 nM.

Example 93-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)benzoicacid

The title compound was prepared as described in General Method B for thepreparation of Example 7 with replacement of methyl6-chloro-1-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylate with ethyl3-bromobenzoate. MS (ESI) m/z: 495.1 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ7.66 (d, J=7.6 Hz, 2H), 7.63-7.54 (m, 1H), 7.45 (br s, 1H), 7.38-7.26(m, 2H), 7.19 (br d, J=7.0 Hz, 1H), 5.85 (s, 1H), 3.30 (br d, J=12.5 Hz,1H), 3.06-2.96 (m, 2H), 2.37-2.25 (m, 3H), 1.72-1.51 (m, 4H), 1.26-1.16(m, 2H), 1.14 (br d, J=2.7 Hz, 2H); FXR EC₅₀=4200 nM.

Example 104-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)benzoicacid

The title compound was prepared as described in General Method B for thepreparation of Example 7 with replacement of methyl6-chloro-1-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylate with ethyl4-bromobenzoate. MS (ESI) m/z: 495.1 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ7.72 (br d, J=8.9 Hz, 2H), 7.68-7.52 (m, 3H), 6.91 (br d, J=8.9 Hz, 2H),5.83 (s, 1H), 3.42 (br d, J=12.8 Hz, 1H), 3.12 (br t, J=9.2 Hz, 2H),2.39-2.24 (m, 3H), 1.63-1.49 (m, 4H), 1.26-1.15 (m, 4H), 1.12 (br s,2H); FXR EC₅₀=135 nM.

2-(3′-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-8-azaspiro[bicyclo[3.2.1]octane-3,1′-cyclobutan]-2′-en-8-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

Step 1. tert-Butyl3′-oxo-8-azaspiro[bicyclo[3.2.1]octane-3,1′-cyclobutane]-8-carboxylate

Zinc-copper couple (28.3 g, 219 mmol) was added to a solution oftert-butyl 3-methylene-8-azabicyclo[3.2.1]octane-8-carboxylate (4.9 g,21.9 mmol) in diethyl ether (43.0 mL). Trichloroacetyl chloride (13.6mL, 121 mmol) in DME (21.5 mL) was added and the reaction mixture wasstirred at room temperature for 36 h. The reaction was carefullyquenched with 1M aqueous K₂HPO₄ (vigorous bubbling) and then filteredthrough Celite (Et₂O wash). The filtrate was concentrated in vacuo anddiluted with MeOH (65.6 mL). Ammonium chloride (4.49 g, 84 mmol) wasadded to the rapidly stirring mixture followed by zinc dust (8.0 g, 122mmol) in two equal portions. After 40 minutes of stirring, the reactionmixture was filtered through Celite (MeOH wash) and concentrated todryness. The residue was taken up in EtOAc and washed with water andbrine. The aqueous layers were back extracted with EtOAc and thecombined organics were dried over Na₂SO₄, filtered and concentrated todryness onto SiO₂. The resulting mixture was purified by flashchromatography on SiO₂ (0-50% EtOAc/hex, Isco 80 g column, ELS detectorused) to give tert-butyl3′-oxo-8-azaspiro[bicyclo[3.2.1]octane-3,1′-cyclobutane]-8-carboxylate(1.1 g, 4.2 mmol, 20% yield) as a white solid. ¹H NMR (400 MHz, CDCl₃) δ4.50-4.13 (m, 2H), 3.12 (d, J=1.8 Hz, 2H), 2.85 (br s, 2H), 2.23-1.87(m, 4H), 1.86-1.66 (m, 4H), 1.48 (s, 9H).

Example 11.2-(3′-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-8-azaspiro[bicyclo[3.2.1]octane-3,1′-cyclobutan]-2′-en-8-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described for the preparation ofExample 1 with replacement of tert-butyl2-oxo-7-azaspiro[3.5]nonane-7-carboxylate with tert-butyl3′-oxo-8-azaspiro[bicyclo[3.2.1]octane-3,1′-cyclobutane]-8-carboxylate.MS (ESI) m/z: 596.5 [M+H]⁺; ˜6:4 Mixture of olefin isomers: ¹H NMR (400MHz, DMSO-d₆) δ 8.21 (dd, J=3.6, 1.4 Hz, 1H), 7.75-7.46 (m, 5H), 6.28(s, 1H), 5.25 (s, 1H), 4.49-4.29 (m, 2H), 2.84 (s, 1H), 2.17-1.89 (m,7H), 1.85-1.59 (m, 3H), 1.31-1.07 (m, 5H); FXR EC₅₀=189 nM.

Example 126-(3′-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-8-azaspiro[bicyclo[3.2.1]octane-3,1′-cyclobutan]-2′-en-8-yl)nicotinicacid

The title compound was prepared as described for the preparation ofExample 11 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl6-fluoronicotinate. MS (ESI) m/z: 522.6 [M+H]⁺; ¹H NMR is for ˜6:4mixture of olefin isomers: ¹H NMR (400 MHz, DMSO-d₆) δ 8.66-8.51 (m,1H), 7.99-7.78 (m, 1H), 7.71-7.48 (m, 3H), 6.71-6.64 (m, 1H), 6.24 (s,1H), 5.18 (s, 1H), 4.57 (br d, J=3.5 Hz, 2H), 2.80 (s, 1H), 2.37-2.23(m, 2H), 2.01-1.82 (m, 5H), 1.83-1.65 (m, 3H), 1.63-1.45 (m, 3H),1.30-1.06 (m, 7H); FXR EC₅₀=182 nM.

Example 132-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]nonan-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

Triethylsilane (70.0 μL, 0.44 mmol) was added to a solution of2-(2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid (Example 1) (10 mg, 0.02 mmol) in TFA (175 μL). The reaction vialwas sealed and heated to 80° C. After 30 minutes the reaction mixturewas concentrated to dryness and the residue was taken up in ˜2 mL of 1:1DMF and MeOH, filtered, and purified via preparative LC/MS with thefollowing conditions: Column: XBridge C18, 19×200 mm, 5-μm particles;Mobile Phase A: 5:95 acetonitrile: water with 10-mM ammonium acetate;Mobile Phase B: 95:5 acetonitrile: water with 10-mM ammonium acetate;Gradient: 30-70% B over 20 minutes, then a 5-minute hold at 100% B;Flow: 20 mL/min. Fractions containing the desired product were combinedand dried via centrifugal evaporation to give2-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]nonan-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid. MS (ESI) m/z: 572.0 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ 8.16 (s,1H), 7.71-7.52 (m, 4H), 3.45-3.34 (m, 1H), 2.28-2.17 (m, 1H), 2.12-1.99(m, 2H), 1.78 (br t, J=10.7 Hz, 2H), 1.74-1.65 (m, 2H), 1.36-1.26 (m,2H), 1.22 (s, 2H), 1.12 (br d, J=7.9 Hz, 3H), 1.08-0.99 (m, 3H); FXREC₅₀=202 nM.

Example 144-(2-(3-(2-Chlorophenyl)-5-cyclopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)benzoicacid

The title compound was obtained during the preparation of Example 10from Pd-mediated dehalogenation during the Buchwald amination step.Alternatively the title compound could be prepared as described forExample 10 with the replacement of 2,6-dichlorobenzaldehyde with2-chlorobenzaldehyde. MS (ESI) m/z: 461.1 [M+H]⁺; ¹H NMR (500 MHz,DMSO-d₆) δ 7.72 (br d, J=8.9 Hz, 2H), 7.66-7.60 (m, 1H), 7.59-7.52 (m,1H), 7.47 (br d, J=4.0 Hz, 2H), 6.92 (br d, J=8.9 Hz, 2H), 5.86 (s, 1H),3.49-3.38 (m, 1H), 3.21-3.06 (m, 2H), 2.92 (q, J=7.1 Hz, 1H), 2.35 (s,2H), 2.32-2.22 (m, 1H), 1.67-1.49 (m, 4H), 1.23 (s, 3H), 1.19-1.12 (m,4H), 1.10 (br d, J=2.4 Hz, 2H); FXR EC₅₀=885 nM.

Example 152-(3′-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-8-azaspiro[bicyclo[3.2.1]octane-3,1′-cyclobutan]-8-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described for the preparation ofExample 13 with replacement of2-(2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid (Example 1) with2-(3′-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-8-azaspiro[bicyclo[3.2.1]octane-3,1′-cyclobutan]-2′-en-8-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid (Example 11). MS (ESI) m/z: 598.0 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆)δ 8.17 (s, 1H), 7.68-7.52 (m, 4H), 4.48-4.12 (m, 1H), 3.89 (s, 1H),3.58-3.34 (m, 1H), 2.23-2.13 (m, 2H), 2.11-2.03 (m, 1H), 2.03-1.86 (m,3H), 1.83-1.72 (m, 1H), 1.72-1.54 (m, 4H), 1.33 (br d, J=13.7 Hz, 1H),1.10 (br d, J=8.2 Hz, 2H), 1.01 (br d, J=2.4 Hz, 2H); FXR EC₅₀=301 nM.

Example 166-(3′-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-8-azaspiro[bicyclo[3.2.1]octane-3,1′-cyclobutan]-8-yl)nicotinicacid

The title compound was prepared as described for the preparation ofExample 15 with replacement of2-(3′-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-8-azaspiro[bicyclo[3.2.1]octane-3,1′-cyclobutan]-2′-en-8-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid (Example 11) with6-(3′-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-8-azaspiro[bicyclo[3.2.1]octane-3,1′-cyclobutan]-2′-en-8-yl)nicotinicacid (Example 12). MS (ESI) m/z: 523.9 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆)δ 8.57 (d, J=1.5 Hz, 1H), 7.86 (dd, J=9.0, 2.0 Hz, 1H), 7.70-7.51 (m,3H), 6.63 (br d, J=8.9 Hz, 1H), 4.66-4.48 (m, 1H), 4.40 (br s, 1H),2.62-2.57 (m, 1H), 2.25-2.12 (m, 1H), 2.12-1.98 (m, 2H), 1.97-1.77 (m,3H), 1.59 (br t, J=10.1 Hz, 4H), 1.45 (br d, J=11.6 Hz, 1H), 1.20 (br d,J=13.4 Hz, 1H), 1.10 (br d, J=8.2 Hz, 2H), 1.01 (br d, J=2.4 Hz, 2H);FXR EC₅₀=646 nM.

Example 172-(2-(5-Cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of 2,6-dichlorobenzaldehydewith 2-(trifluoromethoxy)benzaldehyde. MS (ESI) m/z: 585.9 [M+H]⁺; ¹HNMR (500 MHz, DMSO-d₆) δ 8.18 (br s, 1H), 7.73-7.63 (m, 1H), 7.63-7.46(m, 4H), 5.92 (s, 1H), 3.70 (br d, J=2.2 Hz, 1H), 3.64 (br s, 2H),3.55-3.43 (m, 1H), 2.47-2.37 (m, 2H), 2.37-2.26 (m, 1H), 1.63 (br s,4H), 1.25-1.12 (m, 3H), 1.10 (br s, 2H); FXR EC₅₀=50 nM.

Example 186-(2-(5-Cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)pyridazine-3-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 17 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl6-chloropyridazine-3-carboxylate. MS (ESI) m/z: 513.2 [M+H]⁺; ¹H NMR(500 MHz, DMSO-d₆) δ 7.78 (br d, J=9.4 Hz, 1H), 7.71-7.60 (m, 1H),7.59-7.44 (m, 3H), 7.30 (br s, 1H), 7.36-7.05 (m, 1H), 5.88 (s, 1H),3.55-3.38 (m, 2H), 2.38 (s, 2H), 2.33-2.22 (m, 1H), 1.55 (br s, 4H),1.25-1.10 (m, 4H), 1.06 (br s, 2H), 0.98 (d, J=6.2 Hz, 1H); FXREC₅₀=1500 nM.

Example 195-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)pyrazine-2-carboxylic acid

The title compound was prepared as described in General Method B for thepreparation of Example 7 with replacement of methyl6-chloro-1-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylate with methyl5-bromopyrazine-2-carboxylate. MS (ESI) m/z: 497.2 [M+H]⁺; ¹H NMR (500MHz, DMSO-d₆) δ 8.59 (br s, 1H), 8.28 (br s, 1H), 7.70-7.52 (m, 3H),5.85 (s, 1H), 4.18-3.95 (m, 4H), 3.53-3.35 (m, 2H), 2.33 (br s, 3H),1.55 (br s, 4H), 1.21 (br d, J=5.0 Hz, 3H), 1.11 (br s, 2H), 1.00 (br d,J=6.1 Hz, 1H); FXR EC₅₀=110 nM.

Example 206-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)quinoline-2-carboxylic acid

The title compound was prepared as described in General Method B for thepreparation of Example 7 with replacement of methyl6-chloro-1-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylate with methyl6-bromoquinoline-2-carboxylate. MS (ESI) m/z: 546.3 [M+H]⁺; ¹H NMR (500MHz, CDCl₃) δ 8.19 (d, J=8.5 Hz, 1H), 7.95 (d, J=8.5 Hz, 1H), 7.91 (d,J=9.4 Hz, 1H), 7.72-7.65 (m, 3H), 7.63-7.55 (m, 1H), 7.22 (d, J=2.2 Hz,1H), 5.86 (s, 1H), 3.59-3.46 (m, 2H), 3.20 (br t, J=9.4 Hz, 2H),2.40-2.30 (m, 3H), 1.78-1.58 (m, 4H), 1.29-1.23 (m, 2H), 1.16-1.09 (m,2H); FXR EC₅₀=63 nM; Mouse in vivo (3 mg/kg, @6 h): Cyp7a1=−98%,Fgf15=+30×.

Example 216-(2-(5-Cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)nicotinic acid

The title compound was prepared as described in General Method A for thepreparation of Example 17 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl6-fluoronicotinate. MS (ESI) m/z: 511.9 [M+H]⁺; ¹H NMR (500 MHz,DMSO-d₆) δ 8.54 (d, J=1.8 Hz, 1H), 7.94-7.82 (m, 1H), 7.72-7.60 (m, 1H),7.55-7.41 (m, 3H), 6.80 (br d, J=9.2 Hz, 1H), 5.87 (s, 1H), 3.81 (br d,J=15.3 Hz, 2H), 3.46-3.27 (m, 2H), 2.36 (s, 2H), 2.31-2.20 (m, 1H),1.60-1.42 (m, 4H), 1.23-1.11 (m, 4H), 1.10-1.00 (m, 2H); FXR EC₅₀=770nM.

Example 225-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)pyrimidine-2-carboxylic acid

The title compound was prepared as described in General Method B for thepreparation of Example 7 with replacement of methyl6-chloro-1-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylate with methyl5-bromopyrimidine-2-carboxylate. MS (ESI) m/z: 497.2 [M+H]⁺; ¹H NMR (500MHz, DMSO-d₆) δ 8.46 (br s, 2H), 7.75-7.50 (m, 3H), 5.85 (s, 1H), 3.68(br d, J=13.7 Hz, 1H), 3.51 (br s, 1H), 3.21 (br s, 2H), 2.32 (s, 3H),1.71-1.51 (m, 4H), 1.27-1.16 (m, 2H), 1.12 (br s, 2H); FXR EC₅₀=1500 nM.

Example 235-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)picolinic acid

The title compound was prepared as described in General Method B for thepreparation of Example 7 with replacement of methyl6-chloro-1-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylate with methyl5-bromopicolinate. MS (ESI) m/z: 496.0 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆)δ 8.29 (br s, 1H), 7.83 (d, J=8.9 Hz, 1H), 7.70-7.54 (m, 3H), 7.37-7.26(m, 1H), 5.84 (s, 1H), 3.83-3.68 (m, 1H), 3.49 (br d, J=13.1 Hz, 1H),3.24-3.11 (m, 2H), 2.31 (s, 3H), 1.68-1.50 (m, 4H), 1.27-1.15 (m, 2H),1.11 (br d, J=2.1 Hz, 2H), 1.00 (d, J=6.4 Hz, 1H); FXR EC₅₀=340 nM.

Example 242-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)benzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with ethyl2-bromobenzo[d]thiazole-6-carboxylate. MS (ESI) m/z: 552.0 [M+H]⁺; ¹HNMR (500 MHz, DMSO-d₆) δ 8.32 (s, 1H), 7.83 (br d, J=7.9 Hz, 1H),7.72-7.56 (m, 3H), 7.43 (br d, J=8.2 Hz, 1H), 5.89 (s, 1H), 3.90 (s,1H), 3.71 (br d, J=11.0 Hz, 1H), 3.17 (s, 1H), 2.41-2.29 (m, 3H), 1.64(br s, 4H), 1.30-1.17 (m, 4H), 1.14 (br d, J=2.4 Hz, 2H); FXR EC₅₀=17nM.

Example 252-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-1-methyl-1H-benzo[d]imidazole-5-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl2-bromo-1-methyl-1H-benzo[d]imidazole-5-carboxylate. MS (ESI) m/z: 549.2[M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ 7.94 (s, 1H), 7.78 (br d, J=8.2 Hz,1H), 7.73-7.65 (m, 2H), 7.65-7.57 (m, 1H), 7.45 (br d, J=8.5 Hz, 1H),5.89 (s, 1H), 3.63 (s, 3H), 3.24-3.11 (m, 1H), 2.36 (s, 3H), 1.82-1.72(m, 2H), 1.70 (br s, 2H), 1.28-1.18 (m, 4H), 1.15 (br d, J=2.1 Hz, 2H);FXR EC₅₀=1020 nM.

Example 262-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)benzo[d]oxazole-5-carboxylicacid

Step 1. Methyl 2-bromobenzo[d]oxazole-5-carboxylate

tert-Butyl nitrite (0.28 g, 2.7 mmol) was added slowly to a 0° C.suspension of copper (II) bromide (0.55 g, 2.5 mmol) in acetonitrile(11.3 mL). After 5 minutes methyl 2-aminobenzo[d]oxazole-5-carboxylate(0.43 g, 2.3 mmol) was added and the reaction mixture was brought toroom temperature. After stirring overnight, the mixture was concentratedonto SiO₂ for purification. The residue was purified by flashchromatography on SiO₂ (0-60% EtOAc/hexanes, Isco 40 g column) to givemethyl 2-bromobenzo[d]oxazole-5-carboxylate (0.16 g, 0.60 mmol, 27%yield) as a white solid. ¹H NMR (400 MHz, CDCl₃) δ 8.44-8.35 (m, 1H),8.13 (dd, J=8.6, 1.8 Hz, 1H), 7.58 (d, J=8.6 Hz, 1H), 3.97 (s, 3H).

Example 26.2-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)benzo[d]oxazole-5-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl2-bromobenzo[d]oxazole-5-carboxylate. MS (ESI) m/z: 536.3 [M+H]⁺; ¹H NMR(500 MHz, DMSO-d₆) δ 7.75 (s, 1H), 7.71-7.63 (m, 3H), 7.63-7.57 (m, 1H),7.45 (d, J=8.2 Hz, 1H), 5.89 (s, 1H), 3.82-3.66 (m, 2H), 3.47 (br d,J=8.2 Hz, 1H), 2.40-2.29 (m, 3H), 1.72-1.56 (m, 4H), 1.28-1.18 (m, 3H),1.15 (br d, J=2.4 Hz, 2H); FXR EC₅₀=157 nM.

Example 276-(2-(5-Cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-5-fluoronicotinicacid

The title compound was prepared as described in General Method A for thepreparation of Example 17 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl6-chloro-5-fluoronicotinate. MS (ESI) m/z: 530.0 [M+H]⁺; ¹H NMR (500MHz, DMSO-d₆) δ 8.46 (br s, 1H), 7.78-7.64 (m, 2H), 7.63-7.47 (m, 3H),5.91 (s, 1H), 3.78 (br d, J=13.7 Hz, 2H), 2.41 (s, 2H), 2.35-2.25 (m,1H), 1.73-1.54 (m, 4H), 1.24 (s, 1H), 1.18 (br d, J=7.6 Hz, 2H), 1.11(br s, 2H); FXR EC₅₀=1100 nM.

Example 286-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-5-fluoronicotinicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl6-chloro-5-fluoronicotinate. MS (ESI) m/z: 514.1 [M+H]⁺; ¹H NMR (500MHz, DMSO-d₆) δ 8.45 (br s, 1H), 7.81-7.69 (m, 1H), 7.68-7.63 (m, 2H),7.62-7.56 (m, 1H), 5.86 (s, 1H), 3.76 (br s, 1H), 2.34 (br s, 3H), 1.60(br s, 4H), 1.29-1.17 (m, 3H), 1.14 (br s, 2H) additional peaks werelost due to water suppression in the ¹H NMR experiment; FXR EC₅₀=453 nM.

Example 292-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of 2,6-dichlorobenzaldehydewith 2-(trifluoromethyl)benzaldehyde. MS (ESI) m/z: 570.1 [M+H]⁺; ¹H NMR(400 MHz, DMSO-d₆) δ 8.20 (d, J=1.5 Hz, 1H), 7.96-7.90 (m, 1H),7.87-7.73 (m, 2H), 7.63-7.53 (m, 2H), 5.79 (s, 1H), 3.81-3.63 (m, 3H),2.33 (s, 4H), 1.69-1.54 (m, 4H), 1.23-1.15 (m, 3H), 1.13 (dt, J=5.4, 2.8Hz, 2H); FXR EC₅₀=14 nM.

Example 306-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)nicotinic acid

The title compound was prepared as described in General Method A for thepreparation of Example 29 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl6-fluoronicotinate. MS (ESI) m/z: 496.1 [M+H]⁺; ¹H NMR (400 MHz,DMSO-d₆) δ 8.58 (d, J=2.0 Hz, 1H), 7.96-7.90 (m, 1H), 7.88 (dd, J=9.0,2.4 Hz, 1H), 7.85-7.73 (m, 2H), 7.57 (d, J=7.0 Hz, 1H), 6.83 (d, J=9.2Hz, 1H), 5.76 (s, 1H), 3.86-3.78 (m, 2H), 3.44-3.33 (m, 2H), 2.32-2.24(m, 3H), 1.51 (br t, J=5.5 Hz, 4H), 1.24-1.15 (m, 2H), 1.15-1.08 (m,2H); FXR EC₅₀=110 nM.

Example 31(6-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)nicotinoyl)glycine

Step A. T3P (45.2 μL, 0.08 mmol) and Et₃N (21.4 μL, 0.15 mmol) wereadded to a solution of6-(2-(5-cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)nicotinicacid (Example 30) (19 mg, 0.04 mmol) and methyl 2-aminoacetate, HCl (9.6mg, 0.08 mmol) in DCE (0.19 mL). The reaction mixture was stirred atroom temperature for and the crude reaction mixture was loaded directlyonto a SiO₂ cartridge for purification by flash chromatography on SiO₂(0-100% EtOAc/hex, Isco 4 g column) to give methyl2-(6-(2-(5-cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)nicotinamido)acetate (10 mg,0.018 mmol, 46.0% yield) as a white foam.

Step B. Methyl2-(6-(2-(5-cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)nicotinamido)acetate(10 mg, 0.02 mmol) was dissolved in THF (136 μL), water (27.2 μL), MeOH(13.6 μL) and then lithium hydroxide monohydrate (3.7 mg, 0.09 mmol) wasadded to the mixture. The reaction vessel was sealed and heated to 60°C. After heating for 2 hours the reaction was quenched with 1N HCl andthen concentrated under a stream of nitrogen to minimum volume. Theresidue was taken up in DMF, filtered and the crude material waspurified via preparative LC/MS with the following conditions: Column:XBridge C18, 19×200 mm, 5-μm particles; Mobile Phase A: 5:95acetonitrile: water with 10-mM ammonium acetate; Mobile Phase B: 95:5acetonitrile: water with 10-mM ammonium acetate; Gradient: 20-60% B over19 minutes, then a 5-minute hold at 100% B; Flow: 20 mL/min. Fractionscontaining the desired product were combined and dried via centrifugalevaporation to give(6-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)nicotinoyl)glycine(7.7 mg, 0.01 mmol, 79% yield). MS (ESI) m/z: 552.9 [M+H]⁺; ¹H NMR (500MHz, DMSO-d₆) δ 8.78 (br d, J=1.2 Hz, 1H), 8.70 (br s, 1H), 8.13 (br t,J=6.6 Hz, 2H), 8.05-7.92 (m, 2H), 7.78 (br d, J=7.3 Hz, 1H), 7.04 (br d,J=9.2 Hz, 1H), 5.96 (s, 1H), 4.06 (br d, J=5.5 Hz, 2H), 4.02-3.90 (m,2H), 1.71 (br s, 4H), 1.45 (s, 2H), 1.42-1.35 (m, 2H), 1.33 (br d, J=2.4Hz, 2H), 1.21 (d, J=6.4 Hz, 2H); FXR EC₅₀=5300 nM.

Example 32(2-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carbonyl)glycine

The title compound was prepared as described for the preparation ofExample 31 with replacement of6-(2-(5-cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)nicotinicacid (Example 30) with2-(2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid (Example 29). MS (ESI) m/z: 627.1 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆)δ 8.80 (br s, 1H), 8.11 (s, 1H), 7.93 (br d, J=7.3 Hz, 1H), 7.86-7.72(m, 2H), 7.62 (br d, J=11.9 Hz, 1H), 7.57 (br d, J=7.3 Hz, 1H), 5.78 (s,1H), 3.92 (br s, 1H), 3.69 (br d, J=13.7 Hz, 1H), 2.37-2.25 (m, 3H),1.71-1.55 (m, 4H), 1.27-1.16 (m, 3H), 1.12 (br d, J=2.4 Hz, 2H);additional ¹H NMR peaks were lost due to water suppression in the ¹H NMRexperiment; FXR EC₅₀=1500 nM.

Example 332-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-2-hydroxy-7-azaspiro[3.5]nonan-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

The title compound was obtained as a minor isolate during thepreparation of Example 1 in General Method A and was purified viapreparative LC/MS with the following conditions: Column: XBridge C18,19×200 mm, 5-μm particles; Mobile Phase A: 5:95 acetonitrile: water with10-mM ammonium acetate; Mobile Phase B: 95:5 acetonitrile: water with10-mM ammonium acetate; Gradient: 25-65% B over 19 minutes, then a5-minute hold at 100% B; Flow: 20 mL/min. Fractions containing thedesired product were combined and dried via centrifugal evaporation. MS(ESI) m/z: 588.2 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ 8.18 (br s, 1H),7.67-7.48 (m, 3H), 6.38 (s, 1H), 3.67 (br s, 1H), 3.54 (br s, 1H), 2.90(s, 1H), 2.74 (s, 1H), 2.38 (br d, J=12.8 Hz, 2H), 2.31-2.18 (m, 1H),1.98 (br s, 2H), 1.54 (br s, 2H), 1.18-1.06 (m, 2H), 1.01-0.90 (m, 2H);FXR EC₅₀=4800 nM.

Example 342-(6-(5-cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-2-azaspiro[3.3]hept-5-en-2-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 29 with replacement of tert-butyl2-oxo-7-azaspiro[3.5]nonane-7-carboxylate with tert-butyl6-oxo-2-azaspiro[3.3]heptane-2-carboxylate. MS (ESI) m/z: 542.6 [M+H]⁺;¹H NMR (400 MHz, DMSO-d₆) δ 12.96 (br s, 1H), 8.23 (d, J=1.5 Hz, 1H),7.99-7.89 (m, 1H), 7.87-7.74 (m, 2H), 7.65-7.51 (m, 2H), 5.67 (s, 1H),4.34 (d, J=9.2 Hz, 2H), 4.24 (d, J=9.2 Hz, 2H), 2.83 (s, 2H), 2.37-2.21(m, 1H), 1.17-1.09 (m, 2H), 0.92-0.79 (m, 2H); FXR EC₅₀=400 nM.

Example 356-(6-(5-cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-2-azaspiro[3.3]hept-5-en-2-yl)nicotinicacid

The title compound was prepared as described in General Method A for thepreparation of Example 34 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl6-fluoronicotinate. MS (ESI) m/z: 468.6 [M+H]⁺; ¹H NMR (400 MHz,DMSO-d₆) δ 8.56 (d, J=2.0 Hz, 1H), 7.97-7.87 (m, 2H), 7.86-7.74 (m, 2H),7.59 (d, J=6.6 Hz, 1H), 6.35 (d, J=9.2 Hz, 1H), 5.64 (s, 1H), 4.17 (d,J=9.2 Hz, 2H), 4.06 (d, J=9.2 Hz, 2H), 2.78 (s, 2H), 2.36-2.26 (m, 1H),1.24-1.17 (m, 2H), 1.17-1.09 (m, 2H); FXR EC₅₀=4300 nM.

Example 362-(2-(5-cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]nonan-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described for the preparation ofExample 13 with replacement of2-(2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid (Example 1) with2-(2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid (Example 29). MS (ESI) m/z: 572.4 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆)δ 8.12 (s, 1H), 7.91 (d, J=7.6 Hz, 1H), 7.85-7.72 (m, 2H), 7.60-7.53 (m,3H), 3.90 (s, 1H), 3.39 (br d, J=9.2 Hz, 2H), 2.27-2.14 (m, 1H),2.09-1.96 (m, 2H), 1.91 (s, 2H), 1.77 (br t, J=10.8 Hz, 2H), 1.73-1.62(m, 2H), 1.45-1.34 (m, 2H), 1.17-1.08 (m, 2H), 1.07-0.97 (m, 2H); FXREC₅₀=116 nM.

Example 372-(2-(5-cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]nonan-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described for the preparation ofExample 36 with replacement of2-(2-(5-cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid (Example 29) with6-(2-(5-cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)nicotinicacid (Example 30). MS (ESI) m/z: 498.4 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆)δ 8.55 (br s, 1H), 7.90 (d, J=7.9 Hz, 1H), 7.85 (br s, 1H), 7.82-7.68(m, 2H), 7.55 (d, J=7.3 Hz, 1H), 6.76 (br d, J=8.9 Hz, 1H), 3.65-3.50(m, 1H), 3.46-3.29 (m, 2H), 2.26-2.15 (m, 1H), 2.04-1.93 (m, 2H), 1.90(s, 1H), 1.72 (br t, J=10.8 Hz, 2H), 1.61-1.48 (m, 2H), 1.32-1.20 (m,2H), 1.15-1.08 (m, 2H), 1.04-0.95 (m, 2H); FXR EC₅₀=1400 nM.

General Method C Example 38(+)-2-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-1,3-dioxa-8-azaspiro[4.5]decan-8-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

Step 1. Methyl5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazole-4-carboxylate

To a 50 mL round bottom flask containing methyl3-cyclopropyl-3-oxopropanoate (1.3 g, 8.9 mmol) was added triethylamine(2.5 mL, 17.8 mmol). The resulting clear solution was stirred at roomtemperature for 15 minutes and was cooled in an ice water bath. To thestirring solution was added a solution of2,6-dichloro-N-hydroxybenzimidoyl chloride (2.0 g, 8.9 mmol, synthesisdescribed in General Method A) in EtOH (4 mL) over the space of 10minutes giving a white suspension. After addition, the resultingsuspension was stirred at room temperature overnight. The reactionmixture was concentrated in vacuo and the residue was purified by flashchromatography on SiO₂ (0-10% EtOAc/hexanes, Isco 80 g column) to givemethyl 5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazole-4-carboxylate (2.4g, 7.7 mmol, 87% yield) as a white solid. ¹H NMR (500 MHz, CDCl₃) δ7.45-7.39 (m, 2H), 7.39-7.33 (m, 1H), 3.71 (s, 3H), 2.93 (tt, J=8.5, 5.2Hz, 1H), 1.47-1.40 (m, 2H), 1.34-1.27 (m, 2H).

Step 2. (5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methanol

To a solution of methyl5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazole-4-carboxylate (3.0 g, 9.6mmol) in THF (11.1 mL) at 0° C. was added 1 M diisobutyl aluminumhydride (20.2 mL, 20.2 mmol) in toluene. The reaction mixture was warmedto room temperature and stirred for 2 hours. The reaction was cooled to0° C. and quenched by the addition of MeOH (2 mL) and 1 M aq. HCl (˜75mL). The mixture was then extracted with EtOAc, and the organic layerwas washed with brine. The organic layer was dried over MgSO₄ andconcentrated to give(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methanol (2.5 g, 8.9mmol, 92% yield) as a white solid, which was used without furtherpurification. ¹H NMR (500 MHz, CDCl₃) δ 7.46 (d, J=1.1 Hz, 1H), 7.45 (s,1H), 7.41-7.36 (m, 1H), 4.44 (s, 2H), 2.22 (tt, J=8.5, 5.2 Hz, 1H), 1.42(br s, 1H), 1.35-1.25 (m, 2H), 1.23-1.11 (m, 2H).

Step 3. 5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazole-4-carbaldehyde

To a solution of(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methanol (2.1 g, 7.4mmol) in DCM (37.0 mL) was added a mixture of pyridinium chlorochromate(6.4 g, 29.6 mmol) and finely ground 3 Å molecular sieves (6.1 g). Theresulting mixture was stirred at room temperature for 30 min and thenfiltered through a pad of Celite. The pad was washed with MeOH/DCM. Thefiltrate was evaporated and the residue was purified by flashchromatography on SiO₂ (0-100% EtOAc/hexanes, Isco 80 g column) to give5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazole-4-carbaldehyde (1.9 g, 6.8mmol, 93% yield) as a white solid. ¹H NMR (500 MHz, CDCl₃) δ 9.67 (s,1H), 7.49-7.44 (m, 2H), 7.43-7.37 (m, 1H), 2.82 (tt, J=8.3, 5.2 Hz, 1H),1.52-1.45 (m, 2H), 1.40-1.33 (m, 2H).

Step 4. (±)-tert-Butyl2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-1,3-dioxa-8-azaspiro[4.5]decane-8-carboxylate

4-Methylbenzenesulfonic acid (1.7 mg, 10.0 μmol) followed by tert-butyl4-hydroxy-4-(hydroxymethyl)piperidine-1-carboxylate (23.1 mg, 0.10 mmol)and 100 mg of oven-dried 3 Å molecular sieves were added to a roomtemperature suspension of5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazole-4-carbaldehyde (28.2 mg,0.1 mmol) in toluene (0.5 mL). The resulting suspension was heated to150° C. overnight. The solids were filtered and washed with DCM (˜10mL). The filtrate was concentrated and the residue was purified by flashchromatography on SiO₂ (0-100% EtOAc/DCM, Isco 40 g column) to givetert-butyl2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-1,3-dioxa-8-azaspiro[4.5]decane-8-carboxylate(12.0 mg, 0.02 mmol, 23% yield) as a white solid. MS (ESI) m/z: 495.1[M+H]⁺; ¹H NMR (500 MHz, CDCl₃) δ 7.45-7.37 (m, 2H), 7.36-7.30 (m, 1H),5.95 (s, 1H), 3.76 (br. s., 1H), 3.61 (d, J=8.0 Hz, 2H), 3.47 (d, J=6.9Hz, 1H), 3.14 (br. s., 1H), 2.94 (br. s., 1H), 2.37-2.14 (m, 1H),1.85-1.65 (m, 1H), 1.46 (s, 10H), 1.38-1.23 (m, 3H), 1.22-1.12 (m, 2H),1.04 (br. s., 1H); FXR EC₅₀=4.8 μM.

Step 5.(+)-2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-1,3-dioxa-8-azaspiro[4.5]decane

Trifluoroacetic acid (0.10 mL, 1.2 mmol) was added to a room temperaturesolution of (±)-tert-butyl2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-1,3-dioxa-8-azaspiro[4.5]decane-8-carboxylate(60 mg, 0.12 mmol) in DCM (2 mL). The reaction mixture was stirred atroom temperature overnight. The excess trifluoroacetic acid was removedin vacuo and the residue was partitioned between EtOAc (5 mL) and 1Maqueous K₂HPO₄ (5 mL). The organic layer was dried over Na₂SO₄, filteredand concentrated to dryness. The crude product was used directly in thenext step.

Example 38. (+)-Ethyl2-(2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-1,3-dioxa-8-azaspiro[4.5]decan-8-yl)-4-fluorobenzo[d]thiazole-6-carboxylate

Cesium carbonate (74.2 mg, 0.23 mmol) and ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate (41.5 mg, 0.14 mmol) wereadded to a room temperature solution of2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-1,3-dioxa-8-azaspiro[4.5]decane(36 mg, 0.09 mmol) in N,N-dimethylacetamide (0.26 mL). After 10 minutesof stirring at room temperature the reaction mixture was heated to 50°C. After 3 hours the reaction mixture was partially concentrated and theresidue was purified by flash chromatography on SiO₂ (5-100%EtOAc/hexanes, Isco 24 g column) to yield (+)-ethyl2-(2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-1,3-dioxa-8-azaspiro[4.5]decan-8-yl)-4-fluorobenzo[d]thiazole-6-carboxylate(23 mg, 0.04 mmol, 39% yield) as a white solid. MS (ESI) m/z: 618.0[M+H]⁺; ¹H NMR (500 MHz, CDCl₃) δ 8.10 (d, J=1.4 Hz, 1H), 7.74 (dd,J=11.3, 1.4 Hz, 1H), 7.50-7.40 (m, 2H), 7.39-7.33 (m, 1H), 6.03 (s, 1H),4.40 (q, J=7.1 Hz, 2H), 4.01 (d, J=12.4 Hz, 1H), 3.91 (d, J=11.8 Hz,1H), 3.66 (d, J=8.3 Hz, 1H), 3.59-3.46 (m, 2H), 3.40-3.16 (m, 1H),2.34-2.16 (m, 1H), 1.93 (dd, J=13.8, 2.5 Hz, 1H), 1.81-1.66 (m, 1H),1.64-1.52 (m, 2H), 1.42 (t, J=7.2 Hz, 3H), 1.34 (dd, J=5.0, 2.2 Hz, 2H),1.24-1.05 (m, 2H); FXR EC₅₀=620 nM.

Example 39(+)-2-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-1,3-dioxa-8-azaspiro[4.5]decan-8-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

Aqueous LiOH 1.0 M (130 μL, 0.13 mmol) was added to a room temperaturesolution of ethyl2-(2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-1,3-dioxa-8-azaspiro[4.5]decan-8-yl)-4-fluorobenzo[d]thiazole-6-carboxylate(16 mg, 0.03 mmol, Example 38) in 1:1 MeOH: THF (260 μL). The reactionmixture was stirred at room temperature overnight and then the excesssolvents were removed. Acetic acid was added until ˜pH 5 was achievedand the mixture was extracted with dichloromethane (10 mL). The organiclayer was collected, dried over MgSO₄, filtered and concentrated invacuo. The crude product was triturated with a 5:1 mixture of hexane:DCM to give(+)-2-(2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-1,3-dioxa-8-azaspiro[4.5]decan-8-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid (12.3 mg, 0.02 mmol, 81% yield) as a white solid. MS (ESI) m/z:590.0 [M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δ 8.04 (s, 1H), 7.67 (d, J=11.0Hz, 1H), 7.39-7.31 (m, 2H), 7.30-7.22 (m, 1H), 5.93 (s, 1H), 3.92 (d,J=11.4 Hz, 1H), 3.79 (br. s., 1H), 3.57 (d, J=8.1 Hz, 1H), 3.46 (d,J=8.1 Hz, 2H), 3.19 (t, J=11.4 Hz, 1H), 2.22-2.09 (m, 1H), 1.83 (d,J=13.0 Hz, 1H), 1.59 (td, J=12.6, 4.5 Hz, 1H), 1.53-1.41 (m, 1H),1.31-1.20 (m, 2H), 1.15-0.95 (m, 3H); FXR EC₅₀=230 nM.

Example 40 Ethyl2-((1R,3S,5S)-2′-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-8-azaspiro[bicyclo[3.2.1]octane-3,4′-[1,3]dioxolan]-8-yl)-4-fluorobenzo[d]thiazole-6-carboxylate

The title compound was prepared as described in General Method C for thepreparation of Example 38 with replacement of tert-butyl4-hydroxy-4-(hydroxymethyl) piperidine-1-carboxylate with tert-butyl(1R,3S,5S)-3-hydroxy-3-(hydroxymethyl)-8-azabicyclo[3.2.1]octane-8-carboxylate.MS (ESI) m/z: 644.1 [M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δ 8.13 (d, J=1.54Hz, 1H), 7.76 (dd, J=1.54, 11.22 Hz, 1H), 7.37-7.45 (m, 2H), 7.29-7.35(m, 1H), 5.87 (s, 1H), 4.41 (q, J=7.04 Hz, 4H), 3.98 (d, J=7.70 Hz, 1H),3.54 (d, J=7.70 Hz, 1H), 2.12-2.39 (m, 5H), 2.07 (br d, J=13.20 Hz, 1H),1.78 (br t, J=9.35 Hz, 1H), 1.37-1.49 (m, 4H), 1.22-1.32 (m, 2H), 1.12(dd, J=1.76, 8.36 Hz, 2H); FXR EC₅₀=3400 nM.

Example 412-((1R,5S)-2′-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-8-azaspiro[bicyclo[3.2.1]octane-3,4′-[1,3]dioxolan]-8-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described General Method C for thepreparation of Example 39 with replacement of tert-butyl4-hydroxy-4-(hydroxymethyl) piperidine-1-carboxylate with tert-butyl(1R,3S,5S)-3-hydroxy-3-(hydroxymethyl)-8-azabicyclo[3.2.1]octane-8-carboxylate.MS (ESI) m/z: 616.6 [M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δ 8.19 (s, 1H), 7.80(br d, J=11.00 Hz, 1H), 7.37-7.47 (m, 2H), 7.30-7.36 (m, 1H), 5.71-6.01(m, 1H), 4.29-4.64 (m, 2H), 3.98 (d, J=7.70 Hz, 1H), 3.78 (s, 1H), 3.54(d, J=7.70 Hz, 2H), 1.99-2.39 (m, 7H), 1.88 (s, 1H), 1.78 (s, 1H),1.58-1.68 (m, 1H), 1.41-1.52 (m, 1H), 1.20-1.35 (m, 3H), 1.12 (dd,J=1.65, 8.47 Hz, 2H), 0.82-1.04 (m, 2H); FXR EC₅₀=1700 nM.

Example 422-(2-(5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)-1,3-dioxa-8-azaspiro[4.5]decan-8-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described General Method C for thepreparation of Example 39 with replacement of 2,6-dichlorobenzaldehydewith 2-(trifluoromethoxy)benzaldehyde. MS (ESI) m/z: 606.1 [M+H]⁺; ¹HNMR (500 MHz, DMSO-d₆) δ 8.11 (s, 1H), 7.70-7.62 (m, 1H), 7.59-7.48 (m,4H), 5.94 (s, 1H), 3.84-3.67 (m, 1H), 3.64-3.37 (m, 1H), 3.24-3.15 (m,1H), 2.94-2.89 (m, 1H), 2.46-2.39 (m, 1H), 1.90 (s, 1H), 1.85 (br d,J=13.1 Hz, 1H), 1.75-1.64 (m, 1H), 1.64-1.53 (m, 1H), 1.22-1.05 (m, 7H);FXR EC₅₀=1000 nM.

Example 436-(2-(5-Cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)-1,3-dioxa-8-azaspiro[4.5]decan-8-yl)nicotinicacid

The title compound was prepared as described General Method C for thepreparation of Example 42 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl6-fluoronicotinate. MS (ESI) m/z: 532.4 [M+H]⁺; ¹H NMR (500 MHz,DMSO-d₆) δ 8.58 (s, 1H), 7.92-7.86 (m, 2H), 7.69-7.63 (m, 2H), 7.58-7.48(m, 7H), 6.83 (d, J=9.2 Hz, 4H), 5.94 (s, 6H), 3.96 (br d, J=14.0 Hz,1H), 3.93-3.83 (m, 1H), 3.67-3.50 (m, 1H), 3.41-3.29 (m, 1H), 3.17 (s,1H), 3.16-3.08 (m, 1H), 2.48-2.41 (m, 1H), 1.75 (br d, J=13.1 Hz, 1H),1.61-1.50 (m, 1H), 1.47-1.38 (m, 1H), 1.17 (br d, J=7.0 Hz, 2H),1.14-1.07 (m, 2H); FXR EC₅₀=5300 nM.

Example 446-(2-(5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)-1,3-dioxa-8-azaspiro[4.5]decan-8-yl)picolinicacid

The title compound was prepared as described General Method C for thepreparation of Example 42 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl6-fluoropicolinate. MS (ESI) m/z: 532.4 [M+H]⁺; ¹H NMR (500 MHz,DMSO-d₆) δ 7.72-7.63 (m, 1H), 7.63-7.43 (m, 5H), 7.21 (d, J=7.3 Hz, 1H),6.96 (d, J=8.5 Hz, 1H), 5.93 (s, 1H), 3.90-3.72 (m, 2H), 3.64-3.44 (m,2H), 3.33 (br t, J=9.9 Hz, 1H), 3.13 (br t, J=10.7 Hz, 1H), 1.74 (br d,J=13.4 Hz, 1H), 1.58 (br t, J=9.8 Hz, 1H), 1.50-1.39 (m, 1H), 1.17 (brd, J=7.0 Hz, 4H), 1.14-1.07 (m, 2H); FXR EC₅₀=5000 nM.

Example 452-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-methylbenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl2-chloro-4-methylbenzo[d]thiazole-6-carboxylate. MS (ESI) m/z: 566.1[M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ 8.14 (br s, 1H), 7.67 (br d, J=1.2Hz, 2H), 7.66 (s, 1H), 7.62-7.57 (m, 1H), 5.88 (s, 1H), 3.71 (br d,J=11.9 Hz, 1H), 3.59-3.41 (m, 1H), 2.45 (s, 3H), 2.39-2.29 (m, 3H), 1.91(s, 1H), 1.72-1.55 (m, 4H), 1.30-1.18 (m, 4H), 1.16-1.10 (m, 2H); FXREC₅₀=11 nM.

Example 462-(2-(3-(3-Chloropyridin-2-yl)-5-cyclopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of 2,6-dichlorobenzaldehydewith 3-chloropicolinaldehyde. MS (ESI) m/z: 537.2 [M+H]⁺; ¹H NMR (500MHz, DMSO-d₆) δ 8.69 (d, J=4.6 Hz, 1H), 8.20 (s, 1H), 8.15 (d, J=8.2 Hz,1H), 7.64 (dd, J=8.2, 4.6 Hz, 1H), 7.59 (d, J=11.6 Hz, 1H), 5.91 (s,1H), 3.82-3.64 (m, 1H), 3.56-3.40 (m, 1H), 2.37 (s, 2H), 2.35-2.27 (m,1H), 1.74-1.56 (m, 4H), 1.28-1.17 (m, 2H), 1.17-1.09 (m, 2H); FXREC₅₀=200 nM.

Example 472-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of 2,6-dichlorobenzaldehydewith 3,5-dichloroisonicotinaldehyde. MS (ESI) m/z: 571.1 [M+H]⁺; ¹H NMR(500 MHz, DMSO-d₆) δ 8.86 (s, 2H), 8.17 (s, 1H), 7.58 (br d, J=11.4 Hz,1H), 6.02 (s, 1H), 3.73 (br d, J=13.8 Hz, 2H), 3.66-3.44 (m, 2H), 2.42(s, 2H), 2.40-2.28 (m, 1H), 1.69 (br s, 4H), 1.32-1.20 (m, 4H),1.19-1.10 (m, 2H); FXR EC₅₀=25 nM.

Example 482-(2-(5-Cyclopropyl-3-(2,6-dichloro-4-fluorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of 2,6-dichlorobenzaldehydewith 2,6-dichloro-4-fluorobenzaldehyde. MS (ESI) m/z: 588.1 [M+H]⁺; ¹HNMR (500 MHz, DMSO-d₆) δ 8.22 (s, 1H), 7.78 (d, J=8.5 Hz, 2H), 7.60 (brd, J=11.3 Hz, 1H), 5.96 (s, 1H), 3.74 (br s, 1H), 3.55 (br d, J=8.2 Hz,1H), 3.33-3.14 (m, 1H), 3.04-2.95 (m, 1H), 2.41 (s, 2H), 2.39-2.29 (m,1H), 1.68 (br s, 4H), 1.23 (br d, J=7.9 Hz, 2H), 1.16 (br d, J=2.4 Hz,2H); FXR EC₅₀=115 nM.

2-(2-(3-(2,6-Dichlorophenyl)-5-isopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of cyclopropylacetylene withisopropylacetylene. MS (ESI) m/z: 572.1 [M+H]⁺; ¹H NMR (500 MHz, CDCl₃)δ 8.13 (s, 1H), 7.75 (d, J=11.1 Hz, 1H), 7.46-7.39 (m, 2H), 7.39-7.32(m, 1H), 5.78 (s, 1H), 3.83-3.71 (m, 2H), 3.57 (ddd, J=13.0, 7.9, 4.7Hz, 2H), 3.40-3.29 (m, 1H), 2.63 (s, 1H), 2.36 (s, 2H), 1.84-1.71 (m,4H), 1.45 (d, J=6.9 Hz, 6H); FXR EC₅₀=57 nM.

Example 502-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethoxy)benzo[d]thiazole-6-carboxylicacid

Step 1. Methyl2-amino-4-(trifluoromethoxy)benzo[d]thiazole-6-carboxylate

Bromine (0.22 mL, 4.2 mmol) dissolved in acetic acid (2.8 mL) was addedto a 0° C. solution of methyl 4-amino-3-(trifluoromethoxy)benzoate (1.0g, 4.2 mmol) and sodium thiocyanate (1.4 g, 17.0 mmol) in acetic acid(5.7 mL). The reaction mixture was brought to room temperature andstirred overnight. More bromine (0.22 mL, 4.2 mmol) was added and thereaction mixture was heated to 50° C. After heating through the weekendthe reaction mixture was partitioned between EtOAc and water. Theorganic layer was collected, dried over Na₂SO₄, filtered, andconcentrated in vacuo. The residue was purified by flash chromatographyon SiO₂ (0-100% EtOAc/hexanes, Isco 24 g column) to give methyl2-amino-4-(trifluoromethoxy)benzo[d]thiazole-6-carboxylate (0.21 g, 0.72mmol, 17% yield) as ayellow solid. ¹H NMR (400 MHz, CDCl₃) δ 8.27 (d,J=1.5 Hz, 1H), 7.94 (t, J=1.5 Hz, 1H), 5.85 (br s, 2H), 3.96 (s, 3H).

Step 2. Methyl2-bromo-4-(trifluoromethoxy)benzo[d]thiazole-6-carboxylate

tert-Butyl nitrite (0.11 mL, 0.86 mmol) was added to a rapidly stirringsuspension of copper (II) bromide (0.18 g, 0.79 mmol) in acetonitrile(3.6 mL). After 5 minutes, the resulting dark brown mixture was added toa flask containing methyl2-amino-4-(trifluoromethoxy)benzo[d]thiazole-6-carboxylate (0.21 g, 0.72mmol) suspended in acetonitrile (0.5 mL). The reaction mixture wasstirred at room temperature for 2.5 h and was then diluted with EtOAcand SiO₂ was added. The mixture was concentrated to give a free-flowingsolid that was purified by flash chromatography on SiO₂ (0-40%EtOAc/hexanes, Isco 24 g column) to give methyl2-bromo-4-(trifluoromethoxy)benzo[d]thiazole-6-carboxylate (0.13 g, 0.37mmol, 51% yield) as a white solid. ¹H NMR (400 MHz, CDCl₃) δ 8.50 (d,J=1.5 Hz, 1H), 8.06 (quin, J=1.4 Hz, 1H), 4.01 (s, 3H); ¹⁹F NMR (377MHz, CDCl₃) δ −57.69 (s).

Example 50.2-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethoxy)benzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 47 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl2-bromo-4-(trifluoromethoxy)benzo[d]thiazole-6-carboxylate. MS (ESI)m/z: 637.2 [M+H]⁺; ¹H NMR (500 MHz, CDCl₃) δ 8.62 (s, 2H), 8.24 (s, 1H),7.90 (s, 1H), 5.83 (s, 1H), 3.83-3.74 (m, 2H), 3.62-3.51 (m, 2H), 2.43(s, 2H), 2.22-2.11 (m, 1H), 1.83-1.69 (m, 5H), 1.30 (br d, J=4.6 Hz,2H), 1.18 (br d, J=7.6 Hz, 2H); FXR EC₅₀=11 nM.

Example 512-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-5-methoxybenzo[d]thiazole-6-carboxylicacid

Step 1. Ethyl 2-bromo-5-methoxybenzo[d]thiazole-6-carboxylate

The title compound can be prepared by the two-step procedure describedin Example 50 for the preparation of methyl2-bromo-4-(trifluoromethoxy)benzo[d]thiazole-6-carboxylate with thereplacement of methyl 4-amino-3-(trifluoromethoxy)benzoate with ethyl2-amino-4-methoxybenzoate. ¹H NMR (500 MHz, CDCl₃) δ 8.23 (s, 1H), 7.55(s, 1H), 4.40 (q, J=7.2 Hz, 2H), 3.98 (s, 3H), 1.41 (t, J=7.0 Hz, 3H).

Example 51.2-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-5-methoxybenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 47 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with ethyl2-bromo-5-methoxybenzo[d]thiazole-6-carboxylate. MS (ESI) m/z: 583.1[M+H]⁺; ¹H NMR (500 MHz, CDCl₃) δ 8.62 (s, 2H), 8.36 (s, 1H), 7.13 (s,1H), 5.83 (s, 1H), 4.07 (s, 3H), 3.81-3.68 (m, 3H), 3.60-3.47 (m, 2H),2.59 (s, 2H), 2.43 (s, 2H), 2.20-2.09 (m, 1H), 1.85-1.67 (m, 4H), 1.30(br d, J=4.6 Hz, 2H), 1.18 (br d, J=7.7 Hz, 2H); FXR EC₅₀=72 nM.

Example 522-(2-(3-(3,5-Dichloropyridin-4-yl)-5-isopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 47 with replacement of cyclopropylacetylene withisopropylacetylene. MS (ESI) m/z: 573.3 [M+H]⁺; ¹H NMR (500 MHz, CDCl₃)δ 8.65 (s, 2H), 8.13 (s, 1H), 7.75 (br d, J=11.0 Hz, 1H), 5.83 (s, 1H),3.84-3.75 (m, 2H), 3.66-3.50 (m, 2H), 3.36 (dquin, J=13.8, 6.9 Hz, 1H),2.39 (s, 2H), 1.88-1.69 (m, 4H), 1.46 (br d, J=6.9 Hz, 6H); FXR EC₅₀=58nM.

Example 537-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)cinnoline-3-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 7 with replacement of methyl6-chloro-1-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylate with ethyl7-chlorocinnoline-3-carboxylate, HCl. MS (ESI) m/z: 547.3 [M+H]⁺; ¹H NMR(500 MHz, DMSO-d₆) δ 8.54 (br s, 1H), 8.02 (br d, J=9.16 Hz, 1H), 7.83(br d, J=8.85 Hz, 1H), 7.67-7.75 (m, 2H), 7.59-7.66 (m, 2H), 5.91 (s,1H), 3.69 (br s, 2H), 3.32-3.46 (m, 1H), 3.21 (s, 1H), 2.40 (m, 3H),1.61-1.83 (m, 4H), 1.14-1.36 (m, 4H); FXR EC₅₀=46 nM.

Example 547-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)cinnoline-3-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 53 with replacement of 2,6-dichlorobenzaldehydewith 2-(trifluoromethyl)benzaldehyde. MS (ESI) m/z: 547.2 [M+H]⁺; ¹H NMR(500 MHz, DMSO-d₆) δ 8.52 (s, 1H), 7.99 (d, J=9.16 Hz, 1H), 7.94 (br d,J=7.63 Hz, 1H), 7.81 (br dd, J=7.63, 14.04 Hz, 3H), 7.59 (br d, J=6.41Hz, 2H), 5.72-5.87 (m, 1H), 3.66 (br d, J=13.73 Hz, 2H), 3.33 (br t,J=8.39 Hz, 1H), 3.17 (dd, J=5.49, 10.38 Hz, 1H), 2.33 (m, 3H), 1.65 (brdd, J=3.36, 12.51 Hz, 4H), 1.08-1.30 (m, 4H); FXR EC₅₀=177 nM.

Example 557-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)cinnoline-3-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 53 with replacement of 2,6-dichlorobenzaldehydewith 3,5-dichloroisonicotinaldehyde. MS (ESI) m/z: 548.0 [M+H]⁺; ¹H NMR(500 MHz, Methanol-d₄) δ 8.92 (s, 1H), 8.74 (s, 2H), 8.14 (d, J=9.63 Hz,1H), 7.90-8.07 (m, 1H), 7.15-7.32 (m, 1H), 5.94 (s, 1H), 3.96 (br d,J=13.75 Hz, 2H), 3.53-3.76 (m, 2H), 2.56 (s, 2H), 2.36 (s, 1H), 1.84 (brt, J=4.13 Hz, 4H), 1.19-1.47 (m, 4H); FXR EC₅₀=191 nM.

Example 567-(2-(3-(2-Chloro-6-fluorophenyl)-5-cyclopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)cinnoline-3-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 53 with replacement of 2,6-dichlorobenzaldehydewith 2-chloro-6-fluorobenzaldehyde. MS (ESI) m/z: 531.0 [M+H]⁺; ¹H NMR(500 MHz, DMSO-d₆) δ 8.54 (s, 1H), 8.01 (d, J=9.2 Hz, 1H), 7.82 (d,J=9.8 Hz, 1H), 7.70-7.53 (m, 3H), 7.45 (t, J=8.7 Hz, 1H), 5.93 (s, 1H),3.69 (br s, 2H), 2.42 (s, 2H), 2.35 (br s, 1H), 1.68 (br s, 4H),1.27-1.08 (m, 4H), additional signals missing due to water signalsuppression; FXR EC₅₀=194 nM.

Example 577-(2-(3-(2-Chloro-6-methylphenyl)-5-cyclopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)cinnoline-3-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 53 with replacement of 2,6-dichlorobenzaldehydewith 2-chloro-6-methylbenzaldehyde. MS (ESI) m/z: 527.2 [M+H]⁺; ¹H NMR(500 MHz, DMSO-d₆) δ 8.54 (s, 1H), 8.00 (d, J=9.2 Hz, 1H), 7.86-7.77 (m,1H), 7.59 (s, 1H), 7.49-7.43 (m, 2H), 7.39-7.33 (m, 1H), 5.77 (s, 1H),3.72-3.65 (m, 2H), 3.37-3.23 (m, 2H), 2.43-2.30 (m, 3H), 2.11 (s, 3H),1.71-1.59 (m, 4H), 1.26-1.08 (m, 4H); FXR EC₅₀=227 nM.

Example 586-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(difluoromethoxy)quinoline-2-carboxylicacid

Step 1. Methyl 6-bromo-4-(difluoromethoxy)quinoline-2-carboxylate

To a stirred suspension of Cs₂CO₃ (0.98 g, 3.0 mmol) in DMF (5 mL) at 0°C. was added methyl 6-bromo-4-hydroxyquinoline-2-carboxylate (0.28 g,1.0 mmol) and sodium chlorodifluoroacetate (0.46 g, 3.0 mmol). Thereaction mixture was stirred with heating at 80° C. for 30 minutes.After cooling the reaction mixture to room temperature, water (25 mL)was added, and the resulting suspension was stirred for 1 hour. Thesolid was collected by suction filtration and washed with water (2×5mL). After drying under vacuum overnight, methyl6-bromo-4-(difluoromethoxy)quinoline-2-carboxylate (0.28 g, 0.81 mmol,81% yield) was obtained as a white solid. MS (ESI) m/z: 333.9 [M+H]⁺; ¹HNMR (400 MHz, CDCl₃) δ 8.38 (d, J=2.20 Hz, 1H), 8.16 (d, J=9.02 Hz, 1H),7.91 (dd, J=2.20, 9.24 Hz, 1H), 7.85 (t, J=1.10 Hz, 1H), 6.61-7.17 (m,1H), 4.09 (s, 3H).

Example 58.6-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(difluoromethoxy)quinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 54 with replacement of ethyl7-chlorocinnoline-3-carboxylate, HCl with methyl6-bromo-4-(difluoromethoxy)quinoline-2-carboxylate. MS (ESI) m/z: 612.1[M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δ 7.95 (br d, J=9.24 Hz, 1H), 7.77-7.84(m, 1H), 7.74 (s, 1H), 7.54-7.68 (m, 3H), 7.43 (br d, J=6.60 Hz, 1H),7.25 (br d, J=2.20 Hz, 1H), 6.69-7.12 (m, 1H), 5.64 (s, 1H), 3.45-3.63(m, 2H), 3.27 (ddd, J=4.18, 8.14, 12.54 Hz, 2H), 2.37 (s, 2H), 2.15(ddd, J=3.30, 5.01, 8.42 Hz, 1H), 1.65-1.86 (m, 4H), 1.10-1.33 (m, 4H);FXR EC₅₀=2.3 nM.

Example 592-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-methoxybenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 47 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl2-bromo-4-methoxybenzo[d]thiazole-6-carboxylate. MS (ESI) m/z: 583.1[M+H]⁺; ¹H NMR (400 MHz, Methanol-d₄) δ 8.74 (s, 2H), 7.99 (d, J=1.54Hz, 1H), 7.55 (d, J=1.32 Hz, 1H), 5.90-6.01 (m, 1H), 4.01 (s, 3H),3.72-3.88 (m, 2H), 3.59 (s, 2H), 2.51 (s, 2H), 2.25-2.40 (m, 1H), 1.78(br d, J=4.18 Hz, 4H), 1.12-1.37 (m, 4H); FXR EC₅₀=4.1 nM.

Example 602-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-7-(trifluoromethyl)quinoline-5-carboxylicacid

Step 1. Ethyl 7-(trifluoromethyl)quinoline-5-carboxylate

A solution of 3-amino-5-(trifluoromethyl)benzoic acid (0.51 g, 2.5mmol), glycerol (0.36 mL, 5.0 mmol), and 3-nitrobenzenesulfonic acidsodium salt (1.679 g, 7.46 mmol) in 75% H₂SO₄ (5.9 mL) was heated to100° C. for 1.5 h and then to 140° C. for 1 h. The reaction mixture wascooled to room temperature and then EtOH (10 mL) was added and thereaction mixture was heated to 85° C. overnight. The reaction mixturewas cooled to room temperature and poured into 40 mL of ice water with3.3 g NaOH. 1M K₂HPO₄ was added until the solution reached pH-7. Thesolution was extracted with EtOAc. The organic layer was washed withbrine, dried over Na₂SO₄, filtered, and concentrated in vacuo. Theresidue was purified by flash chromatography on SiO₂ (0-100%EtOAc/hexanes, Isco 24 g column) to give a mixture of ethyl7-(trifluoromethyl) quinoline-5-carboxylate and ethyl5-(trifluoromethyl)quinoline-7-carboxylate (0.34 g, 1.3 mmol, 51% yield)as a beige solid. MS (ESI) m/z: 270.0 [M+H]⁺.

Step 2. 5-(Ethoxycarbonyl)-7-(trifluoromethyl)quinoline 1-oxide

m-Chloroperoxybenzoic acid (0.2 g, 0.87 mmol) was added portion wise toa solution of ethyl 7-(trifluoromethyl)quinoline-5-carboxylate (0.18 g,0.67 mmol) in dichloromethane (5.1 mL). The reaction was stirred at roomtemperature overnight. The solvent volume was reduced by ˜25% and thecrude reaction mixture was loaded directly onto a SiO₂ column forpurification by flash chromatography on SiO₂ (0-10% MeOH/DCM, Isco 24 gcolumn) to give 5-(ethoxycarbonyl)-7-(trifluoromethyl)quinoline 1-oxide(0.20 g, 0.68 mmol, 100% yield) as a yellow solid. ¹H NMR (400 MHz,CDCl₃) δ 9.36 (s, 1H), 8.97 (d, J=9.0 Hz, 1H), 8.64 (d, J=5.5 Hz, 1H),8.53 (d, J=2.0 Hz, 1H), 7.53 (dd, J=9.1, 6.1 Hz, 1H), 4.53 (q, J=7.3 Hz,2H), 1.50 (t, J=7.2 Hz, 3H); ¹⁹F NMR (377 MHz, CDCl₃) δ −62.91 (s).

Step 3. Ethyl 2-chloro-7-(trifluoromethyl)quinoline-5-carboxylate

Phosphorus oxychloride (0.039 mL, 0.42 mmol) followed by DMF (0.014 mL,0.18 mmol) were added to a 0° C. solution of5-(ethoxycarbonyl)-7-(trifluoromethyl) quinoline 1-oxide (0.1 g, 0.35mmol) in dichloromethane (3.5 mL). After 5 minutes the reaction mixturewas brought to room temperature. After stirring at room temp for 30 hthe crude reaction mixture was purified by flash chromatography on SiO₂(0-70% EtOAc/hex, Isco 12 g column, product eluted around 20% EtOAc) togive a mixture of ethyl2-chloro-7-(trifluoromethyl)quinoline-5-carboxylate and ethyl2-chloro-5-(trifluoromethyl)quinoline-7-carboxylate as a white solid.The mixture was used in the next step.

Example 60.2-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-7-(trifluoromethyl)quinoline-5-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 47 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with ethyl2-chloro-7-(trifluoromethyl)quinoline-5-carboxylate. MS (ESI) m/z: 615.1[M+H]⁺; ¹H NMR (500 MHz, CDCl₃) δ 9.06 (br d, J=9.2 Hz, 1H), 8.64 (s,2H), 8.18 (br s, 1H), 8.15 (br s, 1H), 7.15 (br d, J=9.6 Hz, 1H), 5.86(s, 1H), 4.07-3.83 (m, 2H), 3.66-3.49 (m, 2H), 2.45 (s, 2H), 2.30-2.12(m, 1H), 1.73 (br s, 4H), 1.38-1.26 (m, 2H), 1.20 (br d, J=7.9 Hz, 2H);FXR EC₅₀=7.2 nM.

Example 612-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-methylbenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 47 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl2-bromo-4-methylbenzo[d]thiazole-6-carboxylate. MS (ESI) m/z: 566.9[M+H]⁺; ¹H NMR (400 MHz, Methanol-d₄) δ 8.64 (s, 2H), 8.09 (d, J=1.10Hz, 1H), 7.74 (d, J=0.66 Hz, 1H), 7.67 (s, 1H), 5.67-6.00 (m, 1H), 3.76(s, 2H), 3.54 (br d, J=8.36 Hz, 2H), 2.49 (s, 3H), 2.45 (s, 2H), 2.25(s, 1H), 2.00 (s, 1H), 1.64-1.89 (m, 4H), 1.05-1.44 (m, 5H); FXREC₅₀=7.2 nM.

Example 622-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)thiazolo[5,4-b]pyridine-5-carboxylicacid

Step 1.4-(7-(5-Chlorothiazolo[5,4-b]pyridin-2-yl)-7-azaspiro[3.5]non-1-en-2-yl)-5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazole

A suspension of5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)-4-(7-azaspiro[3.5]non-1-en-2-yl)isoxazole(150 mg, 0.40 mmol), 2-bromo-5-chlorothiazolo[5,4-b]pyridine (119 mg,0.48 mmol), and cesium carbonate (325 mg, 1.0 mmol) in DMA (1.2 mL) washeated at 50° C. for 3 hours. The crude reaction mixture was purifieddirectly by flash chromatography on SiO₂ (0-100% EtOAc/hexanes, Isco 40g column) to yield4-(7-(5-chlorothiazolo[5,4-b]pyridin-2-yl)-7-azaspiro[3.5]non-1-en-2-yl)-5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazole(166 mg, 0.29 mmol, 73% yield) as a gum. MS (ESI) m/z: 543.9 [M+H]⁺; ¹HNMR (400 MHz, CDCl₃) δ 8.63 (s, 2H), 7.60 (d, J=8.36 Hz, 1H), 7.19 (d,J=8.36 Hz, 1H), 5.81 (s, 1H), 3.73 (td, J=5.06, 13.42 Hz, 2H), 3.49(ddd, J=4.62, 8.03, 13.09 Hz, 2H), 2.44 (s, 2H), 2.10-2.25 (m, 1H),1.70-1.81 (m, 4H), 1.13-1.36 (m, 4H).

Step 2. Methyl2-(2-(5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)thiazolo[5,4-b]pyridine-5-carboxylate

A mixture of4-(7-(5-chlorothiazolo[5,4-b]pyridin-2-yl)-7-azaspiro[3.5]non-1-en-2-yl)-5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazole(100 mg, 0.18 mmol), methanol (5 mL, 0.18 mmol),1,3-bis(diphenylphosphanyl)propane (9.1 mg, 0.022 mmol), palladium(II)acetate (4.9 mg, 0.022 mmol), and potassium carbonate (40.6 mg, 0.29mmol) in DMF (2.5 mL) was heated under CO (48 psi) in a pressure bottleat 85° C. for one day. The mixture was diluted with ethyl acetate (10mL) and filtered through Celite. The filtrate was concentrated undervacuum to dryness. The residue was dissolved in ethyl acetate (20 mL)and washed with water (10 mL). The organic layer was dried overanhydrous Na₂SO₄, filtered and concentrated to dryness in vacuo. Theresidue was purified by flash chromatography on SiO₂ (0-100%EtOAc/hexanes, Isco 40 g column) to yield methyl2-(2-(5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)thiazolo[5,4-b]pyridine-5-carboxylatewith approximately 85% purity. The material was used for next stepwithout further purification. MS (ESI) m/z: 568.0 [M+H]⁺.

Example 62.2-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)thiazolo[5,4-b]pyridine-5-carboxylicacid

To a solution of methyl2-(2-(5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)thiazolo[5,4-b]pyridine-5-carboxylate(20 mg, 0.04 mmol) in 1:1 methanol/THF (0.35 mL), was added 1N NaOH(0.11 mL, 0.11 mmol). The reaction mixture was heated to 60° C. for 15minutes. The crude reaction mixture was acidified with TFA and purifieddirectly by C-18 reverse phase flash chromatography (10-100% B in A, A=10:90:0.1 MeCN:H₂O:TFA, B=90:10:0.1 MeCN:H₂O:TFA, 18 min lineargradient, Isco 12 g C-18 gold column) to yield2-(2-(5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)thiazolo[5,4-b]pyridine-5-carboxylicacid (11 mg, 0.019 mmol, 54% yield) as a red solid. MS (ESI) m/z: 554.0[M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δ 8.67 (s, 2H), 8.17 (d, J=8.36 Hz, 1H),7.83 (s, 1H), 5.74-5.95 (m, 1H), 3.75-3.98 (m, 2H), 3.48-3.69 (m, 2H),2.48 (s, 2H), 2.11-2.26 (m, 1H), 1.82 (br t, J=4.95 Hz, 4H), 1.14-1.51(m, 4H); FXR EC₅₀=24 nM.

Example 632-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)benzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 47 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with ethyl2-chlorobenzo[d]thiazole-6-carboxylate. MS (ESI) m/z: 552.9 [M+H]⁺; ¹HNMR (400 MHz, Methanol-d₄) δ 8.67 (s, 2H), 8.31 (d, J=1.54 Hz, 1H), 7.99(dd, J=1.76, 8.58 Hz, 1H), 7.69 (s, 1H), 7.46 (d, J=8.58 Hz, 1H),5.76-5.99 (m, 1H), 3.79 (br d, J=13.64 Hz, 2H), 3.48-3.68 (m, 2H), 2.50(s, 2H), 2.15-2.34 (m, 1H), 2.04 (s, 1H), 1.70-1.89 (m, 4H), 1.16-1.40(m, 4H); FXR EC₅₀=62 nM.

Example 642-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-8-methoxyquinoline-5-carboxylicacid

Step 1. Methyl 8-methoxyquinoline-5-carboxylate

A solution of 3-amino-4-methoxybenzoic acid (3.3 g, 19.7 mmol), glycerol(2.9 mL, 39.5 mmol), and 3-nitrobenzenesulfonic acid sodium salt (13.3g, 59.2 mmol) in 75% H₂SO₄ (47.0 mL) was heated to 100° C. for 2 h andthen 140° C. for 1 h. The reaction mixture was cooled to roomtemperature and then MeOH (40 mL) was added and the reaction mixture washeated to 60° C. overnight. The reaction mixture was cooled to roomtemperature and poured into ice water and made basic with 12 M NH₄OH.The resulting mixture was extracted with EtOAc. The layers wereseparated and the aqueous layer was further extracted with EtOAc (2×).The combined organic layers were washed with brine, dried over MgSO₄,filtered, and concentrated in vacuo. The crude product was dry loadedonto SiO₂ and purified by flash chromatography on SiO₂ (0-100%EtOAc/hexanes) to provide methyl 8-methoxyquinoline-5-carboxylate (2.2g, 9.9 mmol, 50% yield) as a white solid. ¹H NMR (500 MHz, CDCl₃) δ 9.50(dd, J=8.8, 1.7 Hz, 1H), 9.00 (dd, J=3.9, 1.7 Hz, 1H), 8.37 (d, J=8.3Hz, 1H), 7.59 (dd, J=8.8, 4.1 Hz, 1H), 7.08 (d, J=8.3 Hz, 1H), 4.19 (s,3H), 4.00 (s, 3H).

Step 2. 8-Methoxy-5-(methoxycarbonyl)quinoline 1-oxide

m-Chloroperoxybenzoic acid (0.97 g, 4.3 mmol) was added portion wise toa solution of methyl 8-methoxyquinoline-5-carboxylate (0.72 g, 3.3 mmol)in dichloromethane (25.5 mL). The reaction was stirred at roomtemperature overnight. The solvent volume was reduced by ˜25% and thecrude reaction mixture was loaded directly onto a SiO₂ column forpurification by flash chromatography on SiO₂ (0-10% MeOH/DCM, Isco 40 gcolumn) to give 8-methoxy-5-(methoxycarbonyl)quinoline 1-oxide (0.6 g,2.6 mmol, 78% yield) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 9.02(dd, J=8.9, 1.0 Hz, 1H), 8.47 (dd, J=6.2, 1.1 Hz, 1H), 8.31 (d, J=8.8Hz, 1H), 7.34 (dd, J=8.9, 6.1 Hz, 1H), 7.07 (d, J=8.8 Hz, 1H), 4.11 (s,3H), 3.99 (s, 3H).

Step 3. Methyl 2-chloro-8-methoxyquinoline-5-carboxylate

Phosphorus oxychloride (0.29 mL, 3.1 mmol) followed by DMF (0.10 mL, 1.3mmol) were added to a 0° C. solution of8-methoxy-5-(methoxycarbonyl)quinoline 1-oxide (0.6 g, 2.6 mmol) indichloromethane (26 mL). After 5 minutes the reaction mixture wasbrought to room temperature. After 24 h the crude reaction mixture waspurified by flash chromatography on SiO₂ (0-85% EtOAc/hexanes, followedby 0-10% DCM/MeOH, Isco 40 g column) to give methyl2-chloro-8-methoxyquinoline-5-carboxylate (0.58 g, 2.3 mmol, 90% yield)as a white solid. ¹H NMR (400 MHz, DMSO-d₆) δ 9.35 (d, J=9.0 Hz, 1H),8.34 (d, J=8.4 Hz, 1H), 7.78 (d, J=9.0 Hz, 1H), 7.38 (d, J=8.6 Hz, 1H),4.07 (s, 3H), 3.92 (s, 3H).

Example 64.2-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-8-methoxyquinoline-5-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 47 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl2-chloro-8-methoxyquinoline-5-carboxylate. MS (ESI) m/z: 577.0 [M+H]⁺;¹H NMR (500 MHz, CDCl₃) δ 9.16 (br d, J=9.6 Hz, 1H), 8.61 (s, 2H), 8.08(br d, J=8.3 Hz, 1H), 7.09 (br d, J=9.5 Hz, 1H), 6.92 (br d, J=8.2 Hz,1H), 5.85 (s, 1H), 4.06 (s, 3H), 4.02-3.83 (m, 2H), 3.71 (s, 1H),3.62-3.47 (m, 2H), 2.41 (s, 2H), 2.26-2.11 (m, 1H), 1.73 (br s, 4H),1.29 (br d, J=4.6 Hz, 2H), 1.17 (br d, J=7.9 Hz, 2H); FXR EC₅₀=49 nM.

Example 652-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)quinoline-5-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 47 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl2-chloroquinoline-5-carboxylate. MS (ESI) m/z: 547.2 [M+H]⁺; ¹H NMR (500MHz, DMSO-d₆) δ 8.92-8.86 (m, 1H), 8.85 (s, 2H), 7.79 (br d, J=7.0 Hz,1H), 7.73 (br d, J=7.9 Hz, 1H), 7.57 (br t, J=7.8 Hz, 1H), 7.31 (br d,J=9.5 Hz, 1H), 5.98 (s, 1H), 3.98-3.85 (m, 2H), 3.48 (br d, J=7.3 Hz,1H), 2.39 (br s, 3H), 1.59 (br s, 4H), 1.33-1.19 (m, 3H), 1.16 (br s,2H); FXR EC₅₀=65 nM.

Example 666-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)nicotinicacid

The title compound was prepared as described in General Method A for thepreparation of Example 47 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl6-fluoronicotinate. MS (ESI) m/z: 497.1 [M+H]⁺; ¹H NMR (500 MHz,DMSO-d₆) δ 8.88 (s, 2H), 8.60 (d, J=1.8 Hz, 1H), 7.89 (dd, J=9.2, 2.1Hz, 1H), 6.84 (d, J=9.2 Hz, 1H), 5.98 (s, 1H), 3.93-3.74 (m, 2H),2.44-2.30 (m, 3H), 1.56 (br s, 4H), 1.32-1.20 (m, 4H), 1.17 (br d, J=2.7Hz, 2H); FXR EC₅₀=342 nM.

Example 676-(2-(5-Cyclopropyl-3-(2,6-dichloro-4-fluorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)nicotinicacid

The title compound was prepared as described in General Method A for thepreparation of Example 66 with replacement of3,5-dichloroisonicotinaldehyde with 2,6-dichloro-4-fluorobenzaldehyde.MS (ESI) m/z: 514.1 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ 8.61 (s, 1H),7.91 (br d, J=8.9 Hz, 1H), 7.78 (br d, J=8.2 Hz, 2H), 6.85 (br d, J=9.2Hz, 1H), 5.94 (s, 1H), 3.95-3.80 (m, 2H), 2.44-2.29 (m, 3H), 1.94 (s,2H), 1.57 (br s, 4H), 1.29-1.18 (m, 2H), 1.17 (br s, 2H) additionalsignals lost due to water suppression in ¹H NMR; FXR EC₅₀=493 nM.

2-(2-(5-Cyclopropyl-3-(3,5-difluoropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of 2,6-dichlorobenzaldehydewith 3,5-difluoroisonicotinaldehyde. MS (ESI) m/z: 539.0 [M+H]⁺; ¹H NMR(400 MHz, DMSO-d₆) δ 8.81 (s, 2H), 8.21 (d, J=1.5 Hz, 1H), 7.59 (dd,J=1.5, 11.5 Hz, 1H), 6.15 (s, 1H), 3.88-3.40 (m, 4H), 2.51 (s, 2H),2.40-2.28 (m, 1H), 1.70 (dd, J=4.7, 7.0 Hz, 4H), 1.30-1.06 (m, 4H); FXREC₅₀=442 nM.

Example 692-(2-(5-Cyclopropyl-3-(3-fluoro-5-methoxypyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

The title compound was obtained as an additional isolate during thepreparation of Example 68 by displacement of one fluorine by MeOH. MS(ESI) m/z: 551.0 [M+H]⁺; ¹H NMR (500 MHz, Methanol-d₄) δ 8.40 (s, 1H),8.34 (s, 1H), 8.15 (d, J=1.5 Hz, 1H), 7.66 (dd, J=1.5, 11.6 Hz, 1H),5.99 (s, 1H), 4.00 (s, 3H), 3.84 (dt, J=4.9, 13.7 Hz, 2H), 3.61 (ddd,J=4.2, 8.2, 13.0 Hz, 2H), 2.53 (s, 2H), 2.32 (tt, J=5.4, 8.0 Hz, 1H),1.79 (dt, J=5.0, 10.2 Hz, 4H), 1.22 (ddd, J=2.5, 6.3, 7.7 Hz, 4H); FXREC₅₀=730 nM.

Example 702-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-6-methoxybenzo[d]thiazole-4-carboxylicacid

Step 1. Methyl 2-amino-6-methoxybenzo[d]thiazole-4-carboxylate

Methyl 2-amino-5-methoxybenzoate (190 mg, 1.0 mmol) was dissolved inacetonitrile (5.2 mL). Ammonium thiocyanate (120 mg, 1.6 mmol) wasadded, followed by benzyltrimethylammonium tribromide (409 mg, 1.0 mmol)after 3.5 h, the reaction mixture was diluted with EtOAc, washed withsaturated NaHCO₃, then brine, dried over Na₂SO₄, filtered, andconcentrated in vacuo. The crude material was purified by flashchromatography on SiO₂ (0-100% EtOAc/hexanes, 17 minute gradient, Isco12 g column) to give methyl2-amino-6-methoxybenzo[d]thiazole-4-carboxylate (100 mg, 0.42 mmol, 40%yield). ¹H NMR (400 MHz, CDCl₃) δ 7.51 (d, J=2.6 Hz, 1H), 7.35 (d, J=2.6Hz, 1H), 5.89 (br. s., 2H), 3.99 (s, 3H), 3.88 (s, 3H).

Step 2. Methyl 2-bromo-6-methoxybenzo[d]thiazole-4-carboxylate

Copper (II) bromide (159 mg, 0.71 mmol) and tert-butyl nitrite (85 μL,0.71 mmol) were dissolved in MeCN (1.7 mL) and allowed to stir 10minutes. Methyl 2-amino-6-methoxybenzo[d]thiazole-4-carboxylate (100 mg,0.42 mmol) was dissolved in MeCN (2.5 mL) and the copper solution wasadded. After 2 h, the reaction mixture was diluted with EtOAc, washedwith 1 N HCl, saturated NaHCO₃, then brine, dried over Na₂SO₄, filtered,and concentrated in vacuo to give methyl2-bromo-6-methoxybenzo[d]thiazole-4-carboxylate. The product was usedwithout further purification. ¹H NMR (400 MHz, CDCl₃) δ 7.68 (d, J=2.6Hz, 1H), 7.46 (d, J=2.6 Hz, 1H), 4.05 (s, 3H), 3.93 (s, 3H).

Example 70.2-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-6-methoxybenzo[d]thiazole-4-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 47 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with2-bromo-6-methoxybenzo[d]thiazole-4-carboxylic acid. MS (ESI) m/z: 583.0[M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ 8.88 (s, 2H), 7.74 (d, J=2.7 Hz,1H), 7.39 (d, J=2.7 Hz, 1H), 6.00 (s, 1H), 3.81 (s, 3H), 3.73-3.62 (m,2H), 3.56-3.45 (m, 2H), 2.43 (s, 2H), 2.36 (td, J=4.3, 8.5 Hz, 1H), 1.70(br t, J=6.2 Hz, 4H), 1.28-1.13 (m, 4H); FXR EC₅₀=4400 nM.

Example 712-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-methoxybenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 29 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl2-bromo-4-methoxybenzo[d]thiazole-6-carboxylate. MS (ESI) m/z: 582.2[M+H]⁺; ¹H NMR (400 MHz, Methanol-d₄) δ 7.98 (d, J=1.54 Hz, 1H),7.86-7.93 (m, 1H), 7.75 (br d, J=1.98 Hz, 2H), 7.54 (d, J=1.54 Hz, 1H),7.48-7.53 (m, 1H), 5.74 (s, 1H), 4.00 (s, 3H), 3.70-3.89 (m, 2H), 3.56(br d, J=8.36 Hz, 2H), 2.43 (s, 2H), 2.30 (s, 1H), 1.62-1.83 (m, 4H),1.11-1.30 (m, 4H); FXR EC₅₀=9.4 nM.

Example 726-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-ethoxyquinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 54 with replacement of ethyl7-chlorocinnoline-3-carboxylate, HCl with methyl6-bromo-4-ethoxyquinoline-2-carboxylate. MS (ESI) m/z: 590.1 [M+H]⁺; ¹HNMR (400 MHz, CDCl₃-d) δ 8.57-8.80 (m, 1H), 7.71-7.88 (m, 3H), 7.56-7.68(m, 2H), 7.42-7.48 (m, 1H), 7.38 (d, J=2.64 Hz, 1H), 5.64 (s, 1H), 4.58(d, J=7.04 Hz, 2H), 3.47-3.70 (m, 2H), 3.33 (br dd, J=4.18, 9.02 Hz,2H), 2.38 (s, 2H), 2.09-2.21 (m, 1H), 1.76 (br t, J=4.95 Hz, 4H), 1.67(t, J=7.04 Hz, 3H), 1.28 (dd, J=2.53, 4.95 Hz, 2H), 1.15 (dd, J=2.64,8.36 Hz, 2H); FXR EC₅₀=10 nM.

Example 736-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(difluoromethoxy)quinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 58 with replacement of2-(trifluoromethyl)benzaldehyde with 3,5-dichloroisonicotinaldehyde. MS(ESI) m/z: 613.0 [M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δ 8.63 (s, 2H), 7.95(d, J=9.24 Hz, 1H), 7.75 (s, 1H), 7.59 (dd, J=2.75, 9.57 Hz, 1H), 7.28(d, J=2.64 Hz, 1H), 6.90 (s, 1H), 5.83 (s, 1H), 3.44-3.64 (m, 2H), 3.29(br s, 2H), 2.44 (s, 2H), 2.19 (ddd, J=3.30, 5.01, 8.42 Hz, 1H),1.68-1.93 (m, 5H), 1.31-1.37 (m, 2H), 1.18 (s, 2H); FXR EC₅₀=16 nM.

Example 746-(2-(3-(3-Chloropyridin-4-yl)-5-cyclopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(difluoromethoxy)quinoline-2-carboxylicacid

The title compound was obtained as a minor isolate from the preparationof Example 73. MS (ESI) m/z: 579.1 [M+H]⁺; ¹H NMR (500 MHz, Methanol-d₄)δ 8.78 (s, 1H), 8.65 (d, J=4.95 Hz, 1H), 8.19 (d, J=9.63 Hz, 1H), 7.95(dd, J=2.48, 9.63 Hz, 1H), 7.91 (s, 1H), 7.51-7.59 (m, 2H), 7.36-7.46(m, 1H), 5.97 (s, 1H), 3.65-3.75 (m, 2H), 3.44 (ddd, J=3.71, 8.60, 12.72Hz, 2H), 2.55 (s, 2H), 2.29-2.37 (m, 1H), 1.77-1.89 (m, 4H), 1.17-1.27(m, 4H); FXR EC₅₀=38 nM.

Example 756-(2-(5-Cyclopropyl-3-(4-(trifluoromethyl)pyridin-3-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(difluoromethoxy)quinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 58 with replacement of2-(trifluoromethyl)benzaldehyde with 4-(trifluoromethyl)nicotinaldehyde.MS (ESI) m/z: 613.1 [M+H]⁺; ¹H NMR (400 MHz, Methanol-d₄) δ 8.94-8.82(m, 1H), 8.12-7.98 (m, 2H), 7.84-7.70 (m, 3H), 7.58-7.18 (m, 2H), 5.77(s, 1H), 3.65-3.53 (m, 2H), 3.37-3.23 (m, 2H), 2.46 (s, 2H), 2.32 (tt,J=5.5, 8.0 Hz, 1H), 1.78 (dt, J=4.9, 10.3 Hz, 4H), 1.29-1.15 (m, 4H);FXR EC₅₀=35 nM.

Example 766-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-methoxynicotinicacid

The title compound was prepared as described in General Method A for thepreparation of Example 47 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl6-chloro-4-methoxynicotinate. MS (ESI) m/z: 527.3 [M+H]⁺; ¹H NMR (500MHz, DMSO-d₆) δ 8.83 (s, 2H), 8.32 (s, 1H), 6.33 (s, 1H), 5.94 (s, 1H),3.83-3.70 (m, 6H), 3.48-3.36 (m, 2H), 2.40-2.31 (m, 3H), 1.57 (br s,4H), 1.28-1.19 (m, 2H), 1.16-1.09 (m, 2H); FXR EC₅₀=2149 nM.

Example 776-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)pyridin-3-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(difluoromethoxy)quinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 58 with replacement of2-(trifluoromethyl)benzaldehyde with 2-(trifluoromethyl)nicotinaldehyde.MS (ESI) m/z: 613.2 [M+H]⁺; ¹H NMR (400 MHz, Methanol-d₄) δ 9.02-8.93(m, 1H), 8.77 (s, 1H), 8.09 (d, J=9.5 Hz, 1H), 7.91 (d, J=5.3 Hz, 1H),7.83-7.76 (m, 2H), 7.60-7.20 (m, 2H), 5.78 (s, 1H), 3.72-3.49 (m, 2H),3.38-3.29 (m, 2H), 2.46 (s, 2H), 2.33 (tt, J=5.6, 7.9 Hz, 1H), 1.78 (dt,J=4.8, 10.1 Hz, 4H), 1.35-1.10 (m, 4H); FXR EC₅₀=68 nM.

Example 786-(2-(3-(3-Chloropyridin-2-yl)-5-cyclopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(difluoromethoxy)quinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 58 with replacement of2-(trifluoromethyl)benzaldehyde with 3-chloropicolinaldehyde. MS (ESI)m/z: 579.1 [M+H]⁺; ¹H NMR (400 MHz, Methanol-d₄) δ 8.65 (dd, J=1.4, 4.8Hz, 1H), 8.14 (d, J=9.5 Hz, 1H), 8.09 (dd, J=1.4, 8.3 Hz, 1H), 7.89-7.85(m, 2H), 7.66-7.27 (m, 3H), 5.89 (s, 1H), 3.71-3.60 (m, 2H), 3.44-3.34(m, 2H), 2.46 (s, 2H), 2.34 (tt, J=5.7, 7.6 Hz, 1H), 1.81 (dt, J=4.5,9.5 Hz, 4H), 1.23 (ddq, J=2.4, 5.1, 7.3 Hz, 4H); FXR EC₅₀=121 nM.

Example 797-(2-(5-Cyclopropyl-3-(2,6-difluorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)cinnoline-3-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 53 with replacement of 2,6-dichlorobenzaldehydewith 2,6-difluorobenzaldehyde. MS (ESI) m/z: 515.2 [M+H]⁺; ¹H NMR (500MHz, DMSO-d₆) δ 8.56 (s, 1H), 8.02 (d, J=9.2 Hz, 1H), 7.83 (d, J=9.5 Hz,1H), 7.69 (ddd, J=6.6, 8.4, 15.1 Hz, 1H), 7.61 (s, 1H), 7.33 (t, J=8.0Hz, 2H), 6.00 (s, 1H), 3.76-3.67 (m, 2H), 2.48 (s, 2H), 2.35 (td, J=4.0,8.2 Hz, 1H), 1.76-1.63 (m, 4H), 1.18 (ddt, J=2.7, 5.4, 25.3 Hz, 4H),additional signals missing due to water signal suppression; FXR EC₅₀=733nM.

Example 807-(2-(3-(3-Chloropyridin-4-yl)-5-cyclopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)cinnoline-3-carboxylicacid

The title compound was obtained as a minor isolate during thepreparation of Example 55 from reduction of one chlorine during thePd-catalyzed Buchwald coupling step. MS (ESI) m/z: 514.0 [M+H]⁺; ¹H NMR(500 MHz, Methanol-d₄) δ 8.88 (s, 1H), 8.78 (s, 1H), 8.65 (d, J=4.95 Hz,1H), 8.13 (d, J=9.63 Hz, 1H), 8.00 (s, 2H), 7.55 (d, J=4.95 Hz, 1H),7.26 (d, J=1.93 Hz, 1H), 5.95-6.01 (m, 1H), 3.90-4.09 (m, 2H), 3.67 (brdd, J=5.36, 8.12 Hz, 2H), 3.02 (s, 2H), 2.34 (s, 1H), 1.85 (br s, 4H),1.17-1.41 (m, 4H); FXR EC₅₀=1360 nM.

7-(2-(5-Cyclopropyl-3-(4-(trifluoromethyl)pyridin-3-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)cinnoline-3-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 53 with replacement of 2,6-dichlorobenzaldehydewith 4-(trifluoromethyl)nicotinaldehyde. MS (ESI) m/z: 548.2 [M+H]⁺; ¹HNMR (500 MHz, DMSO-d₆) δ 9.04 (d, J=5.1 Hz, 1H), 8.86 (s, 1H), 8.50 (s,1H), 8.02-7.94 (m, 2H), 7.78 (d, J=9.8 Hz, 1H), 7.57 (s, 1H), 5.84 (s,1H), 3.69-3.61 (m, 2H), 2.39-2.26 (m, 3H), 1.70-1.58 (m, 4H), 1.18 (dt,J=5.1, 38.7 Hz, 4H), additional signals missing due to water signalsuppression; FXR EC₅₀=1540 nM.

Example 826-(2-(5-Cyclopropyl-3-(2,6-difluorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 79 with replacement of ethyl7-chlorocinnoline-3-carboxylate, HCl with ethyl6-chloro-4-(trifluoromethyl)quinoline-2-carboxylate. MS (ESI) m/z: 582.0[M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ 8.21 (s, 1H), 8.08 (d, J=9.5 Hz,1H), 7.88 (d, J=9.5 Hz, 1H), 7.69 (q, J=7.4 Hz, 1H), 7.33 (t, J=8.1 Hz,2H), 7.09 (s, 1H), 6.01 (s, 1H), 3.67-3.59 (m, 2H), 2.48 (s, 2H),2.40-2.30 (m, 1H), 1.77-1.65 (m, 4H), 1.24-1.12 (m, 4H), additionalsignals missing due to water signal suppression; FXR EC₅₀=356 nM.

Example 836-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 82 with replacement of 2,6-difluorobenzaldehydewith 2,6-dichlorobenzaldehyde. MS (ESI) m/z: 614.1 [M+H]⁺; ¹H NMR (500MHz, DMSO-d₆) δ 8.22 (br s, 1H), 8.09 (br s, 1H), 7.79 (br d, J=8.85 Hz,1H), 7.62-7.68 (m, 2H), 7.49-7.62 (m, 2H), 7.06 (br s, 1H), 5.87 (s,1H), 3.55 (br s, 1H), 3.31-3.51 (m, 1H), 2.55 (s, 2H), 2.35 (s, 3H),1.66 (br s, 4H), 1.17-1.25 (m, 2H), 1.13 (br s, 2H); FXR EC₅₀=38 nM.

Example 846-(2-(3-(2,6-Dichlorophenyl)-5-isopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 83 with replacement of cyclopropylacetylene withisopropylacetylene. MS (ESI) m/z: 612.2 [M+H]⁺; ¹H NMR (500 MHz, CDCl₃)δ 8.46-8.30 (m, 1H), 8.06-7.95 (m, 1H), 7.56 (td, J=4.1, 2.6 Hz, 1H),7.44-7.36 (m, 2H), 7.35-7.29 (m, 1H), 7.23-7.17 (m, 1H), 5.77 (s, 1H),3.61-3.46 (m, 2H), 3.40-3.19 (m, 4H), 2.33 (br s, 3H), 1.43 (d, J=6.9Hz, 8H), 1.26 (s, 1H); FXR EC₅₀=127 nM.

Example 856-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylicacid

Step 1. 3,5-Dichloroisonicotinaldehyde oxime

Hydroxylamine hydrochloride (11.8 g, 170 mmol) was added to a roomtemperature solution of 3,5-dichloroisonicotinaldehyde (20 g, 114 mmol)in pyridine (50 mL). After 10 minutes the reaction mixture wasconcentrated in vacuo to remove excess pyridine. The solid was collectedby suction filtration, washed with water and dried in vacuo to give3,5-dichloroisonicotinaldehyde oxime (21.7 g, 114 mmol, 100% yield) as awhite solid. ¹H NMR (400 MHz, DMSO-d₆) δ 12.32 (s, 1H), 8.71 (s, 2H),8.28 (s, 1H).

Step 2. 3,5-Dichloro-N-hydroxyisonicotinimidoyl chloride

3,5-Dichloroisonicotinaldehyde oxime (21.7 g, 114 mmol) was suspended inDMF (114 mL). N-Chlorosuccinimide (16.7 g, 125 mmol) was added in threeportions giving a clear yellow solution. After stirring for 3 hours, thereaction mixture was poured over ice and extracted with Et₂O. Theorganic layer was washed with brine and the combined aqueous layers wereback extracted with Et₂O. The combined organics were dried over Na₂SO₄,filtered and concentrated in vacuo. The residue was purified by flashchromatography on SiO₂ (0-60% EtOAc/hex, Isco 120 g column) to give3,5-dichloro-N-hydroxyisonicotinimidoyl chloride (24.8 g, 110 mmol, 97%yield) as off-white crystals. ¹H NMR (400 MHz, CDCl₃) δ 8.72 (s, 1H),8.62 (s, 2H).

Step 3. 5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazole

Ethynylcyclopropane (12.1 mL, 143 mmol) followed by Et₃N (18.4 mL, 132mmol) were added to a room temperature solution of3,5-dichloro-N-hydroxyisonicotinimidoyl chloride (24.8 g, 110 mmol inDCM (440 mL). After stirring overnight at room temperature, the reactionmixture was concentrated to dryness in vacuo and then taken up inEtOAc/water. The organic layer was washed with brine and the combinedaqueous layers were back extracted with EtOAc. The combined organiclayers were dried over Na₂SO₄, filtered and concentrated in vacuo togive 5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazole (27.3 g, 107mmol, 97% yield as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 8.63 (s,2H), 6.09 (s, 1H), 2.24-2.11 (m, 1H), 1.23-1.07 (m, 4H).

Step 4. 4-Bromo-5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazole

N-Bromosuccinimide (24.8 g, 139 mmol) was added to a room temp solutionof 5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazole (27.3 g, 107mmol) in DMF (143 mL). The reaction mixture was stirred at room tempover the weekend. The reaction mixture was poured over ice and extractedwith Et₂O. The organic layer was washed with brine and the aqueouslayers were back extracted with Et₂O. The combined organic extracts weredried over Na₂SO₄, filtered and concentrated in vacuo to give a yellowsolid. The residue was purified by flash chromatography on SiO₂ (0-30%EtOAc/hex, Isco 220 g column) to give4-bromo-5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazole (29.0 g,74.9 mmol, 81% yield) as a white solid. ¹H NMR (400 MHz, CDCl₃) δ 8.65(s, 2H), 2.19 (tt, J=8.4, 5.1 Hz, 1H), 1.36-1.28 (m, 2H), 1.25-1.17 (m,2H).

Step 5. tert-Butyl2-(5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-2-hydroxy-7-azaspiro[3.5]nonane-7-carboxylate

n-Butyllithium (22.5 mL, 56.1 mmol) was added slowly (over the span of˜30 minutes) to a −78° C. solution of4-bromo-5-cyclopropyl-3-(3,5-dichloropyridin-4-yl) isoxazole (15 g, 44.9mmol) in THF (150 mL). After 10 minutes, tert-butyl2-oxo-7-azaspiro[3.5]nonane-7-carboxylate (10.8 g, 44.9 mmol) as asolution in 8 mL of THF was added slowly to the cold stirring mixture.After 2.5 h the reaction was quenched by the slow addition of 15 mLsaturated aqueous NH₄Cl. The mixture was extracted with EtOAc and theorganic layer was washed with brine. The combined aqueous layers werefurther extracted with EtOAc and the combined organic extracts weredried over Na₂SO₄, filtered and concentrated in vacuo to give an orangesolid residue. The residue was purified by flash chromatography on SiO₂(0-100% EtOAc/DCM, Isco 220 g column, product eluted as a broad lowpeak) to give tert-butyl 2-(5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-2-hydroxy-7-azaspiro[3.5]nonane-7-carboxylate (6.6 g,13.4 mmol, 30% yield) as a tan foam. ¹H NMR (400 MHz, CDCl₃) δ 8.65 (s,2H), 3.34-3.25 (m, 2H), 3.24-3.17 (m, 2H), 2.25-2.11 (m, 4H), 2.09 (s,1H), 1.78-1.65 (m, 2H), 1.49-1.42 (m, 11H), 1.42-1.35 (m, 2H), 1.34-1.28(m, 2H), 1.22-1.11 (m, 2H).

Step 6.5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)-4-(7-azaspiro[3.5]non-1-en-2-yl)isoxazole

Trifluoroacetic acid (5.8 mL, 76 mmol) was added to a flask containingtert-butyl2-(5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-2-hydroxy-7-azaspiro[3.5]nonane-7-carboxylate(3.8 g, 7.6 mmol). After 3 hours the reaction mixture was concentratedto dryness. The residue was taken up in EtOAc and basified withsaturated aqueous K₂C₀₃. The organic layer was washed with brine and thecombined aqueous layers were back extracted with EtOAc. The organicextracts were dried over Na₂SO₄, filtered and concentrated in vacuo togive5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)-4-(7-azaspiro[3.5]non-1-en-2-yl)isoxazoleas a tan foam which was used without further purification.

Step 7. Ethyl6-(2-(5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylate

A slurry of5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)-4-(7-azaspiro[3.5]non-1-en-2-yl)isoxazole(2.0 g, 5.3 mmol), ethyl6-chloro-4-(trifluoromethyl)quinoline-2-carboxylate (1.9 g, 6.4 mmol)and Cs₂CO₃ (3.5 g, 10.6 mmol) in dioxane (35 mL) was degassed bybubbling N₂ through the stirring mixture for 10 minutes.Chloro(2-dicyclohexylphosphino-2′,6′-diisopropoxy-1,1′-biphenyl)[2-(2′-amino-1,1′-biphenyl)]palladium(II)(RuPhos-Pd-G2) (0.20 g, 0.27 mmol) was added and the reaction vessel wassealed and heated to 70° C. After heating overnight the reaction mixturewas diluted with EtOAc, filtered through Celite and concentrated todryness in vacuo. The residue was purified by flash chromatography onSiO₂ (0-100% EtOAc/hex, Isco 80 g column) to give ethyl6-(2-(5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylate(1.2 g, 1.8 mmol, 34% yield) as a yellow solid. ¹H NMR (400 MHz, CDCl₃)δ 8.64 (s, 2H), 8.36 (s, 1H), 8.20 (d, J=9.5 Hz, 1H), 7.60 (dd, J=9.5,2.6 Hz, 1H), 7.22 (s, 1H), 5.84 (s, 1H), 4.56 (q, J=7.2 Hz, 2H),3.63-3.53 (m, 2H), 3.32 (ddd, J=12.8, 8.5, 3.9 Hz, 2H), 2.45 (s, 2H),2.20 (tt, J=8.4, 5.1 Hz, 1H), 1.89-1.72 (m, 4H), 1.49 (t, J=7.2 Hz, 3H),1.36-1.29 (m, 2H), 1.24-1.14 (m, 2H); ¹⁹F NMR (377 MHz, CDCl₃) δ −62.65(s, 3F).

Example 85.6-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylicacid

Ethyl6-(2-(5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylate(1.2 g, 1.8 mmol) was taken up in THF (12 mL), water (4.8 mL), and MeOH(1.2 mL) and then lithium hydroxide (0.43 g, 18.0 mmol) was added to themixture. The reaction was sealed and heated to 50° C. After heating for30 minutes the crude reaction mixture was loaded onto Celite forpurification by C-18 reverse phase flash chromatography (10-100% B in A,A=10:90:0.1 MeCN:H₂O:TFA, B=90:10:0.1 MeCN:H₂O:TFA, 18 min lineargradient, Isco 100 g C-18 gold column) desired fractions were combinedand concentrated to give6-(2-(5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylicacid, Example 85 (1.0 g, 1.6 mmol, 89% yield) as a red solid. MS (ESI)m/z: 615.1 [M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δ 8.66 (s, 2H), 8.45 (s, 1H),8.06 (d, J=9.5 Hz, 1H), 7.65 (dd, J=9.7, 2.6 Hz, 1H), 7.27-7.24 (m, 1H),5.86 (s, 1H), 3.65-3.58 (m, 2H), 3.41-3.32 (m, 2H), 2.47 (s, 2H),2.25-2.16 (m, 1H), 1.89-1.76 (m, 4H), 1.37-1.31 (m, 2H), 1.25-1.17 (m,2H); ¹⁹F NMR (377 MHz, CDCl₃) δ −62.85 (s, 3F); FXR EC₅₀=53 nM; Mouse invivo (3 mg/kg, @ 6 h): Cyp7a1=−94%, Fgf15=+19×.

Example 866-(2-(5-Cyclopropyl-3-(2,6-dichloro-4-fluorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 82 with replacement of 2,6-difluorobenzaldehydewith 2,6-dichloro-4-fluorobenzaldehyde. MS (ESI) m/z: 632.2 [M+H]⁺; ¹HNMR (500 MHz, DMSO-d₆) δ 8.21 (s, 1H), 8.08 (br d, J=9.5 Hz, 1H), 7.88(br d, J=8.5 Hz, 1H), 7.77 (d, J=8.5 Hz, 2H), 7.08 (br s, 1H), 5.94 (s,1H), 3.91 (s, 1H), 3.70-3.56 (m, 2H), 3.36-3.23 (m, 3H), 2.40 (s, 1H),2.38-2.28 (m, 1H), 1.76-1.60 (m, 4H), 1.28-1.18 (m, 2H), 1.16 (br d,J=2.7 Hz, 2H); FXR EC₅₀=68 nM.

Example 876-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-methylquinoline-2-carboxylicacid

The title compound was prepared as described for the preparation ofExample 85 with replacement of ethyl6-chloro-4-(trifluoromethyl)quinoline-2-carboxylate with ethyl6-bromo-4-methylquinoline-2-carboxylate. MS (ESI) m/z: 561.0 [M+H]⁺; ¹HNMR (400 MHz, CDCl₃) δ 8.63 (s, 2H), 7.90-8.14 (m, 2H), 7.57 (s, 1H),7.11 (br s, 1H), 5.79-5.91 (m, 1H), 3.45-3.61 (m, 2H), 3.27 (ddd,J=3.74, 8.58, 12.54 Hz, 2H), 2.69 (s, 3H), 2.44 (s, 2H), 2.19 (s, 1H),1.78-2.03 (m, 4H), 1.27-1.39 (m, 4H); FXR EC₅₀=76 nM.

Example 886-(2-(3-(2-Chloro-6-fluorophenyl)-5-cyclopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 82 with replacement of 2,6-difluorobenzaldehydewith 2-chloro-6-fluorobenzaldehyde. MS (ESI) m/z: 597.9 [M+H]⁺; ¹H NMR(500 MHz, DMSO-d₆) δ 8.21 (s, 1H), 8.07 (d, J=9.4 Hz, 1H), 7.80 (d,J=9.2 Hz, 1H), 7.63 (td, J=6.1, 8.3 Hz, 1H), 7.52 (d, J=8.1 Hz, 1H),7.41 (t, J=8.7 Hz, 1H), 7.09 (s, 1H), 5.94 (s, 1H), 3.63-3.52 (m, 2H),3.34-3.25 (m, 2H), 2.40 (s, 2H), 2.32 (td, J=4.3, 8.5 Hz, 1H), 1.76-1.63(m, 4H), 1.26-1.09 (m, 4H); FXR EC₅₀=150 nM.

Example 896-(2-(3-(2-Chloro-6-methylphenyl)-5-cyclopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 82 with replacement of 2,6-difluorobenzaldehydewith 2-chloro-6-methylbenzaldehyde. MS (ESI) m/z: 594.2 [M+H]⁺; ¹H NMR(500 MHz, DMSO-d₆) δ 8.21 (s, 1H), 8.07 (d, J=9.5 Hz, 1H), 7.83 (dd,J=2.6, 9.5 Hz, 1H), 7.50-7.40 (m, 2H), 7.36 (dd, J=2.9, 5.8 Hz, 1H),7.07 (s, 1H), 5.78 (s, 1H), 3.63-3.55 (m, 2H), 3.31-3.22 (m, 2H),2.42-2.31 (m, 3H), 2.11 (s, 3H), 1.74-1.59 (m, 4H), 1.25-1.10 (m, 4H);FXR EC₅₀=202 nM.

Example 906-(2-(5-Cyclopropyl-3-(4-(trifluoromethyl)pyridin-3-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 82 with replacement of 2,6-difluorobenzaldehydewith 4-(trifluoromethyl)nicotinaldehyde. MS (ESI) m/z: 615.0 [M+H]⁺; ¹HNMR (500 MHz, DMSO-d₆) δ 9.05 (d, J=5.1 Hz, 1H), 8.87 (s, 1H), 8.22 (s,1H), 8.06 (d, J=9.5 Hz, 1H), 7.99 (d, J=5.2 Hz, 1H), 7.82-7.76 (m, 1H),7.07 (s, 1H), 5.85 (s, 1H), 3.58-3.50 (m, 2H), 3.28-3.21 (m, 2H),2.40-2.30 (m, 3H), 1.73-1.61 (m, 4H), 1.28-1.11 (m, 4H); FXR EC₅₀=195nM.

Example 916-(2-(3-(3-Chloropyridin-2-yl)-5-cyclopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 82 with replacement of 2,6-difluorobenzaldehydewith 3-chloropicolinaldehyde. MS (ESI) m/z: 581.2 [M+H]⁺; ¹H NMR (500MHz, DMSO-d₆) δ 8.69 (d, J=4.0 Hz, 1H), 8.20 (s, 1H), 8.15 (d, J=8.2 Hz,1H), 8.07 (br d, J=9.5 Hz, 1H), 7.86 (br d, J=8.5 Hz, 1H), 7.63 (dd,J=8.2, 4.6 Hz, 1H), 7.07 (br s, 1H), 5.89 (s, 1H), 3.70-3.52 (m, 1H),3.31 (br t, J=8.9 Hz, 1H), 2.42-2.27 (m, 3H), 1.75-1.57 (m, 4H),1.27-1.17 (m, 2H), 1.14 (br d, J=2.7 Hz, 2H); FXR EC₅₀=219 nM.

Example 926-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)pyridin-3-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 82 with replacement of 2,6-difluorobenzaldehydewith 2-(trifluoromethyl)nicotinaldehyde. MS (ESI) m/z: 615.3 [M+H]⁺; ¹HNMR (500 MHz, DMSO-d₆) δ 8.93 (dd, J=1.5, 4.8 Hz, 1H), 8.24 (s, 1H),8.20-8.13 (m, 1H), 8.09 (d, J=9.5 Hz, 1H), 7.89 (dd, J=4.7, 7.9 Hz, 1H),7.83-7.72 (m, 1H), 7.06 (s, 1H), 5.83 (s, 1H), 3.56-3.48 (m, 2H),3.26-3.17 (m, 2H), 2.40-2.29 (m, 3H), 1.65 (ddd, J=6.0, 11.1, 18.2 Hz,4H), 1.27-1.08 (m, 4H); FXR EC₅₀=253 nM.

Example 936-(2-(5-Cyclopropyl-3-(3,5-difluoropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 82 with replacement of 2,6-difluorobenzaldehydewith 3,5-difluoroisonicotinaldehyde. MS (ESI) m/z: 583.0 [M+H]⁺; ¹H NMR(400 MHz, DMSO-d₆) δ 8.80 (s, 2H), 8.21 (s, 1H), 8.08 (d, J=9.5 Hz, 1H),7.89 (dd, J=2.6, 9.7 Hz, 1H), 7.08 (d, J=2.4 Hz, 1H), 6.14 (s, 1H),3.66-3.57 (m, 2H), 3.43-3.27 (m, 2H), 2.51 (s, 2H), 2.41-2.30 (m, 1H),1.72 (q, J=4.9, 5.4 Hz, 4H), 1.27-1.10 (m, 4H); FXR EC₅₀=511 nM.

Example 946-(2-(3-(3,5-Dichloropyridin-4-yl)-5-(1-methylcyclopropyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylicacid

The title compound was prepared as described for the preparation ofExample 85 with replacement of cyclopropylacetylene with1-ethynyl-1-methylcyclopropane. MS (ESI) m/z: 629.2 [M+H]⁺; ¹H NMR (500MHz, DMSO-d₆) δ 8.87 (s, 2H), 8.22 (br s, 1H), 8.08 (br d, J=9.2 Hz,1H), 7.85 (br d, J=7.9 Hz, 1H), 7.09 (br s, 1H), 6.07 (s, 1H), 3.67-3.54(m, 1H), 3.39-3.25 (m, 1H), 3.02-2.88 (m, 1H), 2.36 (s, 2H), 1.78-1.61(m, 4H), 1.45 (s, 3H), 1.32-1.21 (m, 2H), 1.18 (br t, J=7.3 Hz, 2H),1.08 (br s, 2H), 0.99 (br s, 2H); FXR EC₅₀=507 nM.

Example 956-(2-(3-(3,5-Dichloropyridin-4-yl)-5-isopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylicacid

The title compound was prepared as described for the preparation ofExample 85 with replacement of cyclopropylacetylene withisopropylacetylene. MS (ESI) m/z: 617.2 [M+H]⁺; ¹H NMR (500 MHz, CDCl₃)δ 8.90-8.52 (m, 2H), 8.42 (br d, J=1.0 Hz, 1H), 8.18-7.93 (m, 1H),7.70-7.45 (m, 1H), 5.83 (s, 1H), 3.56 (br s, 2H), 3.42-3.25 (m, 3H),2.37 (s, 2H), 1.78 (br s, 4H), 1.44 (br d, J=6.9 Hz, 6H), acid OH notobserved, one quinoline C-H under CDCl₃ peak; FXR EC₅₀=37 nM.

Example 966-(2-(3-(2-Chloro-4-fluorophenyl)-5-cyclopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylicacid

The title compound was obtained as a minor isolate during thepreparation of Example 86 by reduction of one chlorine during thePd-catalyzed Buchwald coupling step. MS (ESI) m/z: 598.2 [M+H]⁺; ¹H NMR(500 MHz, DMSO-d₆) δ 8.21 (br s, 1H), 8.08 (br s, 1H), 7.86 (br d, J=9.9Hz, 1H), 7.63 (br d, J=8.8 Hz, 1H), 7.56 (br dd, J=8.3, 6.1 Hz, 1H),7.42-7.31 (m, 1H), 7.11 (br s, 1H), 5.96 (s, 1H), 3.61 (br d, J=13.8 Hz,2H), 3.44-3.24 (m, 1H), 2.43 (s, 1H), 2.37-2.24 (m, 1H), 1.83-1.63 (m,3H), 1.35-1.05 (m, 5H) Additional signals missing due to watersuppression in the ¹H NMR experiment; FXR EC₅₀=869 nM.

Example 976-(2-(5-Cyclopropyl-3-(3-fluoro-5-methoxypyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylicacid

The title compound was obtained as an additional isolate during thepreparation of Example 93 by displacement of one fluorine with MeOHduring the hydrolysis step. MS (ESI) m/z: 595.1 [M+H]⁺; ¹H NMR (400 MHz,Methanol-d₄) δ 8.40 (d, J=3.2 Hz, 1H), 8.34 (t, J=3.2 Hz, 2H), 8.18 (brs, 1H), 7.86-7.77 (m, 1H), 7.25 (s, 1H), 5.98 (d, J=3.3 Hz, 1H), 3.99(s, 3H), 3.72-3.62 (m, 2H), 3.45-3.36 (m, 2H), 2.52 (s, 2H), 2.37-2.28(m, 1H), 1.88-1.72 (m, 4H), 1.21 (q, J=3.5, 6.1 Hz, 4H); FXR EC₅₀=1940nM.

Example 986-(2-(3-Cyclohexyl-5-cyclopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 82 with replacement of 2,6-difluorobenzaldehydewith cyclohexanecarbaldehyde. MS (ESI) m/z: 552.4 [M+H]⁺; ¹H NMR (500MHz, DMSO-d₆) δ 8.23 (s, 1H), 8.10 (d, J=9.3 Hz, 1H), 7.91 (d, J=9.9 Hz,1H), 7.13 (s, 1H), 6.41 (s, 1H), 3.76-3.65 (m, 2H), 2.78-2.67 (m, 3H),2.21 (dq, J=5.3, 8.6 Hz, 1H), 1.97-1.22 (m, 8H), 1.47-1.19 (m, 6H),1.12-0.94 (m, 4H), additional signals missing due to water signalsuppression; FXR EC₅₀=1720 nM.

Example 996-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 82 with replacement of 2,6-difluorobenzaldehydewith 2-(trifluoromethyl)benzaldehyde. MS (ESI) m/z: 614.1 [M+H]⁺; ¹H NMR(500 MHz, Methanol-d₄) δ 8.33 (s, 1H), 8.13 (d, J=9.35 Hz, 1H), 7.89 (d,J=7.15 Hz, 1H), 7.68-7.81 (m, 3H), 7.51 (d, J=7.15 Hz, 1H), 7.21 (br s,1H), 5.67-5.86 (m, 1H), 3.60 (br s, 2H), 3.36 (br d, J=3.58 Hz, 2H),2.42 (s, 2H), 2.31 (s, 1H), 1.66-1.94 (m, 4H), 1.11-1.40 (m, 4H); FXREC₅₀=92 nM.

General Method D Example 1002-(2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)quinoxaline-6-carboxylicacid

Step 1.4-(7-(6-Bromoquinoxalin-2-yl)-7-azaspiro[3.5]non-1-en-2-yl)-5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazole

A slurry of5-cyclopropyl-3-(2,6-dichlorophenyl)-4-(7-azaspiro[3.5]non-1-en-2-yl)isoxazole(25 mg, 0.07 mmol, synthesis described in General Method A),6-bromo-2-chloroquinoxaline (19.5 mg, 0.08 mmol) and Cs₂CO₃ (43.4 mg,0.13 mmol) in dioxane (0.3 mL) was degassed by bubbling nitrogen throughthe mixture for 5 minutes.Chloro(2-dicyclohexylphosphino-2′,6′-diisopropoxy-1,1′-biphenyl)[2-(2′-amino-1,1′-biphenyl)]palladium(II)(RuPhos-Pd-G2) (2.59 mg, 3.33 μmol) was added and the reaction mixturewas sealed and heated to 90° C. for 6 hours. The crude reaction mixturepurified directly by flash chromatography on SiO₂ (0-100% EtOAc/hexanes,Isco 12 g column) to yield4-(7-(6-bromoquinoxalin-2-yl)-7-azaspiro[3.5]non-1-en-2-yl)-5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazole(26 mg, 0.04 mmol, 64% yield) as a gum. ¹H NMR (400 MHz, CDCl₃) δ 8.57(s, 1H), 8.03 (d, J=2.2 Hz, 1H), 7.64 (dd, J=8.8, 2.2 Hz, 1H), 7.53 (d,J=8.8 Hz, 1H), 7.46-7.42 (m, 2H), 7.40-7.34 (m, 1H), 5.80 (s, 1H), 3.94(dt, J=13.5, 5.1 Hz, 2H), 3.66-3.50 (m, 2H), 2.44 (s, 2H), 2.21 (tt,J=8.4, 5.1 Hz, 1H), 1.76 (t, J=5.6 Hz, 4H), 1.36-1.30 (m, 2H), 1.22-1.15(m, 2H).

Step 2.2-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)quinoxaline-6-carbonitrile

A microwave vial containing4-(7-(6-bromoquinoxalin-2-yl)-7-azaspiro[3.5]non-1-en-2-yl)-5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazole(20 mg, 0.03 mmol), Xantphos (4.0 mg, 6.9 μmol), Pd₂(dba)₃ (6.3 mg, 6.9μmol), and zinc cyanide (4.0 mg, 0.03 mmol) was purged three times withnitrogen and anhydrous DMF (0.5 mL) was added. The reaction mixture washeated under microwave irradiation at 110° C. for 1.5 h. The reactionmixture was diluted with EtOAc (10 mL) and washed with brine (10 mL).The organic layer was dried over Na₂SO₄, filtered and concentrated todryness in vacuo. The crude product was purified by flash chromatographyon SiO₂ (0 to 100% EtOAc/hexanes, Isco 12 g column) to yield2-(2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)quinoxaline-6-carbonitrile(11.5 mg, 0.02 mmol, 60% yield) as a gum. ¹H NMR (400 MHz, CDCl₃) δ 8.65(s, 1H), 8.19 (d, J=1.8 Hz, 1H), 7.74-7.68 (m, 1H), 7.67-7.62 (m, 1H),7.48-7.42 (m, 2H), 7.40-7.34 (m, 1H), 5.79 (s, 1H), 4.02 (dt, J=13.5,5.0 Hz, 2H), 3.68-3.59 (m, 2H), 2.46 (s, 2H), 2.20 (tt, J=8.4, 5.1 Hz,1H), 1.77 (t, J=5.6 Hz, 4H), 1.37-1.31 (m, 2H), 1.24-1.15 (m, 2H).

Example 100.2-(2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)quinoxaline-6-carboxylicacid

To a solution of2-(2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)quinoxaline-6-carbonitrile(11.5 mg, 0.02 mmol) in EtOH (72.5 μL), was added NaOH (21.8 μL, 0.11mmol). The reaction mixture was sealed and heated to 90° C. for 2 h. Thecrude reaction mixture was purified by C-18 reverse phase flashchromatography (10-100% B in A, A=10:90:0.1 MeCN:H₂O:TFA, B=90:10:0.1MeCN:H₂O:TFA, 18 min linear gradient, Isco 24 g C-18 gold column)desired fractions were combined and concentrated to give2-(2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)quinoxaline-6-carboxylicacid (9.6 mg, 0.017 mmol, 79% yield) as a yellow solid. MS (ESI) m/z:546.9 [M+H]⁺; ¹H NMR (400 MHz, Methanol-d₄) δ 8.69 (s, 1H), 8.44 (d,J=1.98 Hz, 1H), 8.11 (dd, J=1.87, 8.69 Hz, 1H), 7.59 (d, J=8.58 Hz, 1H),7.37-7.52 (m, 3H), 5.80 (s, 1H), 4.03 (td, J=4.98, 13.59 Hz, 2H),3.52-3.73 (m, 2H), 2.43 (s, 2H), 2.18-2.32 (m, 1H), 1.72 (br t, J=5.50Hz, 4H), 1.07-1.40 (m, 6H); FXR EC₅₀=33 nM.

Example 1012-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)quinoxaline-6-carboxylicacid

The title compound was prepared as described for the preparation ofExample 100 with replacement of 2,6-dichlorobenzaldehyde with2-(trifluoromethyl)benzaldehyde. MS (ESI) m/z: 547.0 [M+H]⁺; ¹H NMR (400MHz, Methanol-d₄) δ 8.81-8.91 (m, 1H), 8.50 (d, J=1.76 Hz, 1H), 8.20(dd, J=1.98, 8.80 Hz, 1H), 7.85-7.99 (m, 1H), 7.71-7.80 (m, 2H), 7.68(d, J=8.80 Hz, 1H), 7.46-7.57 (m, 1H), 5.75 (s, 1H), 3.96-4.35 (m, 2H),3.60-3.89 (m, 2H), 2.46 (s, 1H), 2.31 (s, 1H), 1.63-1.90 (m, 4H),1.10-1.44 (m, 4H); FXR EC₅₀=17 nM.

Example 1022-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)thiazolo[5,4-b]pyridine-5-carboxylicacid

The title compound was prepared as described for the preparation ofExample 62 with replacement of 3,5-dichloroisonicotinaldehyde with2-(trifluoromethyl)benzaldehyde. MS (ESI) m/z: 553.2 [M+H]⁺; ¹H NMR (500MHz, CDCl₃) δ 8.15 (br d, J=8.25 Hz, 1H), 7.83 (br d, J=7.43 Hz, 1H),7.76 (br d, J=8.25 Hz, 1H), 7.62-7.73 (m, 2H), 7.46 (br d, J=6.60 Hz,1H), 5.57-5.82 (m, 1H), 3.78 (br d, J=11.55 Hz, 2H), 3.46-3.65 (m, 2H),2.42 (s, 2H), 2.15 (br d, J=4.68 Hz, 1H), 1.24-1.38 (m, 4H), 1.18 (m,4H); FXR EC₅₀=18 nM.

Example 1032-(2-(3-(2-Chloro-6-methylphenyl)-5-cyclopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of 2,6-dichlorobenzaldehydewith 2-chloro-6-methylbenzaldehyde. MS (ESI) m/z: 549.9 [M+H]⁺; ¹H NMR(500 MHz, DMSO-d₆) δ 8.21 (s, 1H), 7.60 (d, J=11.5 Hz, 1H), 7.48-7.42(m, 2H), 7.40-7.33 (m, 1H), 5.80 (s, 1H), 3.78-3.69 (m, 2H), 3.53-3.45(m, 2H), 2.43-2.29 (m, 3H), 2.12 (s, 3H), 1.66 (br s, 4H), 1.24-1.10 (m,4H); FXR EC₅₀=19 nM.

Example 1046-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-isopropoxyquinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 99 with replacement of ethyl6-chloro-4-(trifluoromethyl) quinoline-2-carboxylate with methyl6-bromo-4-isopropoxyquinoline-2-carboxylate. MS (ESI) m/z: 604.2 [M+H]⁺;¹H NMR (500 MHz, Methanol-d₄) δ 8.08-8.36 (m, 1H), 7.83-8.03 (m, 2H),7.64-7.82 (m, 3H), 7.51 (br d, J=6.60 Hz, 1H), 7.43 (br s, 1H),5.63-5.84 (m, 1H), 5.20-5.46 (m, 1H), 3.62 (br s, 4H), 2.42 (br s, 2H),2.29 (br d, J=4.40 Hz, 1H), 1.75 (br s, 4H), 1.60 (br s, 6H), 1.20 (brs, 4H); FXR EC₅₀=59 nM.

Example 1056-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-methoxyquinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 99 with replacement of ethyl6-chloro-4-(trifluoromethyl) quinoline-2-carboxylate with methyl6-bromo-4-methoxyquinoline-2-carboxylate. MS (ESI) m/z: 576.1 [M+H]⁺; ¹HNMR (400 MHz, Methanol-d₄) δ 8.10 (d, J=9.46 Hz, 1H), 7.86-7.93 (m, 1H),7.72-7.85 (m, 3H), 7.71 (s, 1H), 7.47-7.60 (m, 1H), 7.43 (d, J=2.64 Hz,1H), 5.61-5.86 (m, 1H), 4.31 (s, 3H), 3.51-3.68 (m, 2H), 3.21-3.31 (m,2H), 2.41 (s, 2H), 2.23-2.36 (m, 1H), 1.72-1.89 (m, 4H), 1.11-1.22 (m,4H); FXR EC₅₀=62 nM.

Example 1066-(2-(5-Cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-isopropoxyquinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 104 with replacement of2-(trifluoromethyl)benzaldehyde with 2-(trifluoromethoxy)benzaldehyde.MS (ESI) m/z: 620.2 [M+H]⁺; ¹H NMR (500 MHz, CDCl₃) δ 8.65 (br d, J=9.63Hz, 1H), 7.83 (br d, J=7.70 Hz, 1H), 7.77 (s, 1H), 7.54-7.59 (m, 1H),7.52 (dd, J=1.38, 7.70 Hz, 1H), 7.42 (br d, J=7.15 Hz, 3H), 5.87 (s,1H), 5.16-5.27 (m, 1H), 3.53-3.66 (m, 2H), 3.39 (td, J=3.78, 12.52 Hz,2H), 2.52 (s, 2H), 2.19 (br t, J=4.95 Hz, 1H), 1.79-1.93 (m, 4H), 1.62(d, J=6.05 Hz, 6H), 1.30 (dd, J=2.34, 4.81 Hz, 2H), 1.17 (dd, J=2.61,8.39 Hz, 2H); FXR EC₅₀=21 nM.

Example 1076-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-ethoxyquinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 72 with replacement of2-(trifluoromethyl)benzaldehyde with 3,5-dichloroisonicotinaldehyde. MS(ESI) m/z: 591.1 [M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δ 8.68 (br d, J=9.46Hz, 1H), 8.65 (s, 1H), 7.79-7.87 (m, 1H), 7.76 (s, 1H), 7.39 (d, J=2.42Hz, 1H), 5.84 (s, 1H), 4.59 (d, J=7.04 Hz, 2H), 3.57 (br s, 2H), 3.35(br s, 2H), 2.45 (s, 2H), 2.14-2.27 (m, 1H), 1.82 (br s, 4H), 1.67 (t,J=7.04 Hz, 3H), 1.06-1.50 (m, 4H); FXR EC₅₀=31 nM.

Example 1086-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-methoxyquinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 107 with replacement of methyl6-bromo-4-ethoxyquinoline-2-carboxylate with methyl6-bromo-4-methoxyquinoline-2-carboxylate. MS (ESI) m/z: 577.0 [M+H]⁺; ¹HNMR (400 MHz, Methanol-d₄) δ 8.68 (s, 2H), 8.11 (d, J=9.46 Hz, 1H), 7.78(dd, J=2.53, 9.57 Hz, 1H), 7.72 (d, J=2.64 Hz, 1H), 7.44 (d, J=2.42 Hz,1H), 5.89 (s, 1H), 4.30 (s, 3H), 3.57 (br s, 2H), 3.22-3.32 (m, 2H),2.48 (s, 2H), 2.13-2.36 (m, 1H), 1.81 (br d, J=4.18 Hz, 4H), 1.17-1.34(m, 4H); FXR EC₅₀=32 nM; Mouse in vivo (3 mg/kg, @ 6 h): Cyp7a1=−97%,Fgf15=+14×.

Example 1096-(2-(3-(3-Chloropyridin-4-yl)-5-cyclopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-methoxyquinoline-2-carboxylicacid

The title compound was obtained as a minor isolate during thepreparation of Example 108 by reduction of one chlorine during thePd-catalyzed Buchwald coupling step. MS (ESI) m/z: 543.1 [M+H]⁺; ¹H NMR(400 MHz, CDCl₃) δ 8.65 (s, 2H), 8.03 (d, J=9.24 Hz, 1H), 7.61 (s, 2H),7.40 (d, J=2.64 Hz, 1H), 5.86 (s, 1H), 4.16 (s, 3H), 3.44-3.59 (m, 2H),3.12-3.37 (m, 2H), 2.45 (s, 2H), 1.82 (br d, J=2.42 Hz, 7H), 1.13-1.44(m, 8H), 0.91 (s, 3H); FXR EC₅₀=140 nM.

Example 1106-(2-(5-Cyclopropyl-3-(2,6-dichloro-4-fluorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-isopropoxyquinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 104 with replacement of2-(trifluoromethyl)benzaldehyde with 2,6-dichloro-4-fluorobenzaldehyde.MS (ESI) m/z: 622.0 [M+H]⁺; ¹H NMR (500 MHz, CDCl₃) δ 7.96 (br d, J=8.9Hz, 1H), 7.53 (br s, 2H), 7.43-7.27 (m, 1H), 7.24-7.16 (m, 2H), 5.84 (s,1H), 5.02-4.88 (m, 1H), 3.46 (br s, 3H), 3.36-3.15 (m, 2H), 2.40 (s,2H), 2.21-2.12 (m, 1H), 1.81 (br d, J=10.2 Hz, 4H), 1.50 (br s, 6H),1.28 (br s, 2H), 1.14 (br d, J=7.4 Hz, 2H); FXR EC₅₀=37 nM.

Example 1116-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-isopropoxyquinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 104 with replacement of2-(trifluoromethyl)benzaldehyde with 2,6-dichloro-4-fluorobenzaldehyde.MS (ESI) m/z: 605.1 [M+H]⁺; ¹H NMR (500 MHz, Methanol-d₄) δ 8.73 (s,2H), 8.18 (d, J=9.63 Hz, 1H), 7.98 (br d, J=2.75 Hz, 1H), 7.77 (s, 1H),7.46 (d, J=2.75 Hz, 1H), 5.87-6.03 (m, 1H), 5.28-5.46 (m, 1H), 3.56-3.78(m, 2H), 3.40 (td, J=4.47, 8.67 Hz, 2H), 2.51 (s, 2H), 2.25-2.36 (m,1H), 1.71-1.94 (m, 4H), 1.61 (d, J=6.05 Hz, 6H), 1.03-1.34 (m, 4H); FXREC₅₀=47 nM.

Example 1126-(2-(5-Cyclopropyl-3-(2,6-dichloro-4-fluorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-methoxyquinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 108 with replacement of3,5-dichloroisonicotinaldehyde with 2,6-dichloro-4-fluorobenzaldehyde.MS (ESI) m/z: 594.2 [M+H]⁺; ¹H NMR (500 MHz, CDCl₃) δ 7.96 (br d, J=9.2Hz, 1H), 7.58 (s, 1H), 7.54 (br d, J=9.1 Hz, 1H), 7.40 (br s, 1H), 7.21(br d, J=8.0 Hz, 2H), 5.85 (s, 1H), 4.14 (s, 3H), 3.74 (s, 1H),3.61-3.39 (m, 2H), 3.39-3.19 (m, 2H), 2.42 (s, 2H), 2.18 (br d, J=4.6Hz, 1H), 1.83 (br d, J=12.4 Hz, 4H), 1.30 (br d, J=3.7 Hz, 2H), 1.17 (brd, J=7.0 Hz, 2H); FXR EC₅₀=54 nM.

Example 1136-(2-(5-Cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-ethoxyquinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 72 with replacement of2-(trifluoromethyl)benzaldehyde with 2-(trifluoromethoxy)benzaldehyde.MS (ESI) m/z: 606.2 [M+H]⁺; ¹H NMR (500 MHz, CDCl₃) δ 8.68 (br d, J=9.35Hz, 1H), 7.72-8.01 (m, 2H), 7.47-7.66 (m, 2H), 7.35-7.44 (m, 3H),5.70-5.99 (m, 1H), 4.60 (q, J=6.97 Hz, 2H), 3.53-3.77 (m, 3H), 3.37-3.43(m, 2H), 2.51 (s, 2H), 2.13-2.23 (m, 1H), 1.78-1.88 (m, 4H), 1.66-1.78(m, 3H), 1.25-1.35 (m, 2H), 1.11-1.23 (m, 2H); FXR EC₅₀=116 nM.

Example 1142-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)benzo[d]oxazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl2-chlorobenzo[d]oxazole-6-carboxylate. MS (ESI) m/z: 536.1 [M+H]⁺; ¹HNMR (500 MHz, DMSO-d₆) δ 7.78-7.92 (m, 2H), 7.63-7.67 (m, 2H), 7.53-7.63(m, 1H), 7.28 (d, J=8.16 Hz, 1H), 5.91 (s, 1H), 3.65-3.81 (m, 2H),3.46-3.57 (m, 2H), 2.37 (s, 2H), 2.23-2.36 (m, 1H), 1.59-1.71 (m, 4H),1.11-1.29 (m, 4H); FXR EC₅₀=127 nM.

Example 1156-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-methylnicotinicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl6-fluoro-4-methylnicotinate. MS (ESI) m/z: 510.0 [M+H]⁺; ¹H NMR (500MHz, DMSO-d₆) δ 8.55 (s, 1H), 7.63-7.70 (m, 2H), 7.61 (s, 1H), 6.65 (s,1H), 5.71-6.02 (m, 1H), 3.81 (br d, J=13.43 Hz, 2H), 3.26-3.40 (m, 2H),2.43 (s, 3H), 2.32 (s, 3H), 1.52 (br s, 4H), 1.08-1.34 (m, 4H); FXREC₅₀=132 nM.

Example 1166-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)nicotinicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl6-chloro-4-(trifluoromethyl)nicotinate. MS (ESI) m/z: 564.2 [M+H]⁺; ¹HNMR (500 MHz, DMSO-d₆) δ 8.63 (s, 1H), 7.64-7.68 (m, 2H), 7.60 (dd,J=7.02, 8.85 Hz, 1H), 7.03 (s, 1H), 5.77-6.14 (m, 1H), 3.88 (br d,J=13.43 Hz, 2H), 2.55 (s, 3H), 2.35 (s, 2H), 1.56 (br s, 4H), 1.21 (brd, J=8.24 Hz, 2H), 1.11-1.17 (m, 2H); FXR EC₅₀=135 nM.

Example 1176-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-1-methyl-1H-pyrrolo[3,2-c]pyridine-3-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 7 with replacement of methyl6-chloro-1-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylate with methyl6-chloro-1-methyl-1H-pyrrolo[3,2-c]pyridine-3-carboxylate. MS (ESI) m/z:549.1 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ 8.64 (s, 1H), 8.00 (s, 1H),7.64-7.70 (m, 2H), 7.60 (br d, J=7.02 Hz, 1H), 7.14-7.34 (m, 1H),6.97-7.09 (m, 1H), 5.74-5.99 (m, 1H), 3.75 (s, 3H), 3.67 (br s, 1H),3.29 (br s, 1H), 2.89 (s, 1H), 2.73 (s, 1H), 2.35 (s, 3H), 1.64 (br d,J=16.48 Hz, 4H), 1.05-1.36 (m, 4H); FXR EC₅₀=139 nM.

Example 1182-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)pyrimidine-5-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl2-chloropyrimidine-5-carboxylate. MS (ESI) m/z: 497.1 [M+H]⁺; ¹H NMR(500 MHz, DMSO-d₆) δ 8.73 (s, 2H), 7.64-7.71 (m, 2H), 7.62 (s, 1H),5.80-5.92 (m, 1H), 3.94-4.27 (m, 2H), 2.35 (m, 3H), 1.54 (br s, 4H),1.08-1.30 (m, 4H), additional signals missing due to water signalsuppression; FXR EC₅₀=171 nM.

Example 1192-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)pyrimidine-5-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with ethyl2-chloro-4-(trifluoromethyl)pyrimidine-5-carboxylate. MS (ESI) m/z:565.2 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ 8.80 (s, 1H), 7.64 (s, 2H),7.60 (br d, J=7.02 Hz, 1H), 5.79-5.92 (m, 1H), 4.01 (br s, 2H), 2.34 (m,3H), 1.55 (m, 4H), 1.09-1.30 (m, 4H), additional signals missing due towater signal suppression; FXR EC₅₀=316 nM.

Example 1202-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-6-methylpyrimidine-4-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl2-chloro-6-methylpyrimidine-4-carboxylate. MS (ESI) m/z: 511.4 [M+H]⁺;¹H NMR (500 MHz, DMSO-d₆) δ 7.63-7.71 (m, 2H), 7.60 (br d, J=7.32 Hz,1H), 6.84 (s, 1H), 5.84-5.90 (m, 1H), 3.99 (br d, J=12.82 Hz, 2H), 2.32(m, 3H), 2.29 (s, 3H), 1.50 (br s, 4H), 1.07-1.26 (m, 4H), additionalsignals missing due to water signal suppression; FXR EC₅₀=1390 nM.

Example 1216-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-5-methylnicotinicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl6-fluoro-5-methylnicotinate. MS (ESI) m/z: 510.2 [M+H]⁺; ¹H NMR (500MHz, DMSO-d₆) δ 8.55 (br s, 1H), 7.87 (br s, 1H), 7.52-7.76 (m, 3H),5.74-5.94 (m, 1H), 3.08-3.45 (m, 2H), 3.02 (m, 2H), 2.32 (m, 3H), 2.23(s, 3H), 1.52-1.85 (m, 4H), 1.18-1.35 (m, 4H); FXR EC₅₀=2590 nM.

Example 1222-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)quinoline-6-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 7 with replacement of methyl6-chloro-1-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylate with methyl2-chloroquinoline-6-carboxylate. MS (ESI) m/z: 546.1 [M+H]⁺; ¹H NMR (500MHz, Methanol-d₄) δ 8.52-8.61 (m, 1H), 8.45 (d, J=9.63 Hz, 1H), 8.38(dd, J=1.51, 8.67 Hz, 1H), 7.93 (d, J=8.80 Hz, 1H), 7.55-7.62 (m, 3H),7.50-7.55 (m, 1H), 5.87 (s, 1H), 4.11 (br d, J=14.03 Hz, 2H), 3.65-3.90(m, 2H), 2.56 (s, 2H), 2.34 (s, 1H), 2.06 (s, 1H), 1.90 (br t, J=4.26Hz, 4H), 1.14-1.33 (m, 4H); FXR EC₅₀=40 nM.

2-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)pyrrolo[2,1-f][1,2,4]triazine-5-carboxylicacid

Step 1. Ethyl 2-chloropyrrolo[2,1-f][1,2,4]triazine-5-carboxylate

A mixture of ethyl2,4-dichloropyrrolo[2,1-f][1,2,4]triazine-5-carboxylate (1.3 g, 3.4mmol), and sodium acetate (3.8 g, 45.9 mmol) in a mixture of EtOAc (80mL) and 2-propanol (16 mL) was stirred at room temperature underhydrogen atmosphere (balloon pressure). After 2.5 h, the resultingmixture was filtered through a pad of Celite, and the filtrate wasevaporated under reduced pressure. The crude residue was purified byflash chromatography on SiO₂ (0-100% EtOAc/hexanes, Isco 80 g column) togive ethyl 2-chloropyrrolo[2,1-f][1,2,4]triazine-5-carboxylate (470 mg,2.1 mmol, 61% yield) as a yellow solid. MS (ESI) m/z: 226.1 [M+H]⁺; ¹HNMR (600 MHz, CDCl₃) δ 9.46 (s, 1H), 7.77 (d, J=2.6 Hz, 1H), 7.45 (d,J=2.8 Hz, 1H), 4.43 (d, J=7.2 Hz, 2H), 1.44 (t, J=7.1 Hz, 3H).

Example 123.2-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)pyrrolo[2,1-f][1,2,4]triazine-5-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with ethyl2-chloropyrrolo[2,1-f][1,2,4]triazine-5-carboxylate. MS (ESI) m/z: 535.9[M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ 9.19 (br s, 1H), 7.57-7.69 (m, 4H),7.04 (br s, 1H), 5.87 (s, 1H), 3.75-3.96 (m, 2H), 2.34 (m, 3H), 1.56 (brs, 4H), 1.17-1.27 (m, 2H), 1.09-1.17 (m, 2H), additional signals missingdue to water signal suppression; FXR EC₅₀=40 nM.

6-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-2-methylnicotinicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl6-chloro-2-methylnicotinate. MS (ESI) m/z: 510.1 [M+H]⁺; ¹H NMR (500MHz, DMSO-d₆) δ 7.90 (br d, J=9.00 Hz, 1H), 7.64-7.72 (m, 2H), 7.54-7.63(m, 1H), 6.65 (br d, J=8.92 Hz, 1H), 5.76-5.97 (m, 1H), 3.78-3.96 (m,2H), 3.34 (br s, 1H), 2.55 (m, 3H), 2.33 (br s, 3H), 1.52 (br s, 4H),1.10-1.25 (m, 4H), additional signals missing due to water signalsuppression; FXR EC₅₀=78 nM.

Example 1256-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-1-methyl-1H-indazole-3-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 7 with replacement of methyl6-chloro-1-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylate with methyl6-bromo-1-methyl-3a,7a-dihydro-1H-indazole-3-carboxylate. MS (ESI) m/z:549.0 [M+H]⁺; ¹H NMR (500 MHz, Methanol-d₄) δ 8.19-8.31 (m, 1H), 7.95(d, J=1.38 Hz, 1H), 7.53-7.70 (m, 3H), 7.49 (d, J=9.35 Hz, 1H), 5.97 (s,1H), 4.53 (s, 3H), 3.52-3.85 (m, 5H), 2.61 (s, 2H), 2.36 (s, 1H),1.97-2.22 (m, 5H), 1.09-1.51 (m, 5H); FXR EC₅₀=96 nM.

Example 1266-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-3-(trifluoromethyl)imidazo[1,5-a]pyridine-1-carboxylicacid

Step 1. Ethyl 2-(5-bromopyridin-2-yl)-2-(hydroxyimino)acetate

Sodium nitrite (28.3 mg, 0.41 mmol) in water (0.5 mL) was added to themixture of ethyl 2-(5-bromopyridin-2-yl)acetate (100 mg, 0.41 mmol) inAcOH (0.5 mL) at 0° C. The reaction mixture was stirred at roomtemperature for 1 h and water (0.5 mL) was added. Stirring wasmaintained for 1 h and the reaction mixture was basified with 1M aqueousK₂HPO₄, to pH 8-9. The aqueous layer was extracted with EtOAc, and theorganic layer was dried over MgSO₄, filtered, and concentrated in vacuo.The crude product was used directly in next step.

Step 2. Ethyl6-bromo-3-(trifluoromethyl)imidazo[1,5-a]pyridine-1-carboxylate

Ethyl 2-(5-bromopyridin-2-yl)-2-(hydroxyimino)acetate (4.5 g, 16.5 mmol)was suspended in THF (50 mL). TFA (6.2 mL) was added followed by portionwise addition of zinc dust (2.2 g, 33.0 mmol). Trifluoroacetic anhydride(4.7 mL, 33.0 mmol) was added and the reaction mixture was stirred for 1hour. The mixture was filtered through Celite and concentrated in vacuo.Pyridine (25 mL) was added to the residue followed by slow addition oftrifluoroacetic anhydride (4.7 mL, 33.0 mmol). After 1 h the reactionmixture was concentrated in vacuo and purified by flash chromatographyon SiO₂ (0-100% EtOAc/hexanes, Isco 80 g column) to give ethyl6-bromo-3-(trifluoromethyl) imidazo[1,5-a]pyridine-1-carboxylate (5 g,14.8 mmol, 90% yield) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 8.38(s, 1H), 8.24 (dd, J=9.7, 0.9 Hz, 1H), 7.36 (dd, J=9.6, 1.4 Hz, 1H),4.50 (q, J=7.3 Hz, 2H), 1.46 (t, J=7.2 Hz, 3H).

Example 126.6-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-3-(trifluoromethyl)imidazo[1,5-a]pyridine-1-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 7 with replacement of methyl6-chloro-1-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylate with ethyl6-bromo-3-(trifluoromethyl)imidazo[1,5-a]pyridine-1-carboxylate. MS(ESI) m/z: 603.2 [M+H]⁺; ¹H NMR (500 MHz, Methanol-d₄) δ 8.11-8.18 (m,1H), 7.61 (s, 1H), 7.50-7.59 (m, 3H), 7.46 (dd, J=1.79, 10.04 Hz, 1H),5.85 (s, 1H), 3.06 (ddd, J=3.30, 8.60, 12.04 Hz, 2H), 2.43 (s, 2H), 2.33(s, 1H), 2.06 (s, 1H), 1.59-1.91 (m, 4H), 1.05-1.37 (m, 5H); FXREC₅₀=102 nM.

Example 1276-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-3-methylimidazo[1,5-a]pyridine-1-carboxylicacid

Step 1. Methyl 6-bromo-3-methylimidazo[1,5-a]pyridine-1-carboxylate

Potassium carbonate (0.14 g, 0.98 mmol) was added to a solution of6-bromo-3-methylimidazo[1,5-a]pyridine-1-carboxylic acid (0.1 g, 0.39mmol) in DMF (0.78 mL). After 5 minutes, iodomethane (0.04 mL, 0.59mmol) was added to the thick slurry and the reaction mixture was stirredat room temperature overnight. The reaction mixture was diluted withEt₂O and water. The organic layer was washed with brine and the combinedaqueous layers were back extracted with Et₂O. The combined organicextracts were dried over Na₂SO₄, filtered and concentrated in vacuo togive white crystals of a suitable purity to carry on to the next step.¹H NMR (400 MHz, CDCl₃) δ 8.08 (dd, J=9.6, 1.0 Hz, 1H), 8.00-7.97 (m,1H), 7.14 (dd, J=9.7, 1.5 Hz, 1H), 3.98 (s, 3H), 2.70 (s, 3H).

Example 127.6-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-3-methylimidazo[1,5-a]pyridine-1-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 126 with replacement of ethyl6-bromo-3-(trifluoromethyl) imidazo[1,5-a]pyridine-1-carboxylate withmethyl 6-bromo-3-methylimidazo[1,5-a]pyridine-1-carboxylate. MS (ESI)m/z: 549.2 [M+H]⁺; ¹H NMR (500 MHz, Methanol-d₄) δ 7.96-8.14 (m, 1H),7.41-7.70 (m, 5H), 5.86 (s, 1H), 3.35-3.49 (m, 2H), 3.13 (ddd, J=3.58,8.73, 12.17 Hz, 2H), 2.86 (s, 3H), 2.44 (s, 2H), 2.25-2.37 (m, 1H), 2.06(s, 2H), 1.66-1.90 (m, 4H); FXR EC₅₀=267 nM.

Example 1286-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-1-methyl-1H-indole-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 7 with replacement of methyl6-chloro-1-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylate with methyl6-bromo-1-methyl-1H-indole-2-carboxylate. MS (ESI) m/z: 548.2 [M+H]⁺; ¹HNMR (500 MHz, Methanol-d₄) δ 8.10-8.23 (m, 1H), 8.04 (d, J=8.53 Hz, 1H),7.93 (d, J=9.35 Hz, 1H), 7.44-7.67 (m, 3H), 7.18 (d, J=2.20 Hz, 1H),5.85 (s, 1H), 4.87 (s, 3H), 3.48 (br dd, J=5.09, 11.97 Hz, 2H), 3.33 (brs, 2H), 3.04-3.25 (m, 2H), 2.42 (s, 2H), 2.18-2.35 (m, 1H), 1.61-1.97(m, 4H), 1.04-1.36 (m, 4H); FXR EC₅₀=287 nM.

Example 1297-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)isoquinoline-3-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 7 with replacement of methyl6-chloro-1-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylate with methyl7-chloroisoquinoline-3-carboxylate. MS (ESI) m/z: 546.3 [M+H]⁺; ¹H NMR(400 MHz, CDCl₃) δ 9.19-8.84 (m, 1H), 8.66-8.34 (m, 1H), 8.05-7.69 (m,1H), 7.66-7.50 (m, 1H), 7.49-7.29 (m, 3H), 5.78 (s, 1H), 3.52 (br d,J=2.6 Hz, 3H), 3.39-3.21 (m, 2H), 2.99 (s, 1H), 2.47-2.32 (m, 2H),2.26-2.10 (m, 1H), 1.78 (br s, 4H), 1.38-1.23 (m, 2H), 1.22-1.09 (m,2H); FXR EC₅₀=341 nM.

Example 1302-((7-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)quinoxalin-2-yl)oxy)aceticacid

Step 1. tert-Butyl 2-((7-bromoquinoxalin-2-yl)oxy)acetate

Potassium carbonate (0.10 g, 0.75 mmol) was added to a room temperaturesolution of 7-bromoquinoxalin-2-ol (11 g, 0.5 mmol) in acetone (5 mL).After 5 minutes, tert-butyl 2-bromoacetate (0.15 g, 0.75 mmol) wasadded, and the reaction mixture was stirred at 25° C. overnight.Saturated aqueous NH₄C₁ (20 mL) was added and the resulting mixture wasextracted with Et₂O (3×10 mL). The combined organic layers were driedover MgSO₄, filtered, and concentrated in vacuo. The crude product waspurified by flash chromatography on SiO₂ (0-100% EtOAc/hexanes, Isco 40g column) to yield tert-butyl 2-((7-bromoquinoxalin-2-yl)oxy)acetate (26mg, 0.07 mmol, 15% yield). MS (ESI) m/z: 340.9 [M+H]⁺; ¹H NMR (400 MHz,CDCl₃) δ 8.48-8.67 (m, 1H), 7.98 (d, J=1.98 Hz, 1H), 7.91 (d, J=8.80 Hz,1H), 7.68 (dd, J=2.20, 8.80 Hz, 1H), 4.94 (s, 2H), 1.51 (s, 9H).

Example 130.2-((7-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)quinoxalin-2-yl)oxy)aceticacid

The title compound was prepared as described in General Method B for thepreparation of Example 7 with replacement of methyl6-chloro-1-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylate withtert-butyl 2-((7-bromoquinoxalin-2-yl)oxy)acetate and hydrolysis of thetert-butyl ester with TFA instead of LiOH. MS (ESI) m/z: 577.0 [M+H]⁺;¹H NMR (400 MHz, Methanol-d₄) δ 8.37 (s, 1H), 7.90 (d, J=9.02 Hz, 1H),7.48-7.61 (m, 4H), 7.37 (d, J=2.64 Hz, 1H), 5.88 (s, 1H), 5.08 (s, 2H),3.50-3.65 (m, 2H), 3.39 (dt, J=4.29, 8.53 Hz, 2H), 2.48 (s, 2H), 2.34(s, 1H), 1.76-1.96 (m, 4H), 1.16-1.30 (m, 4H); FXR EC₅₀=373 nM.

Example 1316-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indole-3-carboxylicacid

Step 1. Methyl6-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indole-3-carboxylate

Sodium hydride (48.0 mg, 1.2 mmol) was added to a room temp solution ofmethyl 6-bromo-1H-indole-3-carboxylate (0.25 g, 1.0 mmol) in THF (50mL). After stirring for 15 minutes, SEM-C₁ (0.21 mL, 1.2 mmol) was addedand the resulting suspension was stirred at 25° C. overnight. Water (50mL) was added to the reaction mixture giving a sticky precipitate. Thesolvent was decanted and the residue was purified by flashchromatography on SiO₂ (0-30% EtOAc/hexanes, Isco 24 g column) to yieldmethyl6-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indole-3-carboxylate(0.36 g, 0.83 mmol) as a gum. MS (ESI) m/z: 384.0 [M+H]⁺; ¹H NMR (500MHz, CDCl₃) δ 8.06 (d, J=8.53 Hz, 1H), 7.86 (s, 1H), 7.72 (d, J=1.65 Hz,1H), 7.43 (dd, J=1.65, 8.53 Hz, 1H), 5.48 (s, 2H), 4.80 (s, 1H), 3.94(s, 3H), 1.29 (br t, J=7.15 Hz, 3H), −0.23-0.22 (m, 11H).

Example 131.6-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indole-3-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 7 with replacement of methyl6-chloro-1-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylate with methyl6-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indole-3-carboxylate. MS(ESI) m/z: 664.2 [M+H]⁺; ¹H NMR (500 MHz, Methanol-d₄) δ 8.10-8.23 (m,1H), 8.04 (d, J=8.53 Hz, 1H), 7.93 (d, J=9.35 Hz, 1H), 7.44-7.67 (m,3H), 7.18 (d, J=2.20 Hz, 1H), 5.85 (s, 1H), 4.87 (s, 3H), 3.48 (br dd,J=5.09, 11.97 Hz, 2H), 3.33 (br s, 2H), 3.04-3.25 (m, 2H), 2.42 (s, 2H),2.18-2.35 (m, 1H), 1.61-1.97 (m, 4H), 1.04-1.36 (m, 4H); FXR EC₅₀=445nM.

Example 1327-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-8-methylquinoline-3-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 7 with replacement of methyl6-chloro-1-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylate with methyl7-bromo-8-methylquinoline-3-carboxylate. MS (ESI) m/z: 560.0 [M+H]⁺; ¹HNMR (500 MHz, Methanol-d₄) δ 9.11-9.45 (m, 2H), 8.14 (d, J=9.08 Hz, 1H),7.76 (d, J=9.08 Hz, 1H), 7.42-7.65 (m, 3H), 5.83-6.00 (m, 1H), 3.35-3.45(m, 2H), 3.10-3.30 (m, 2H), 2.66 (s, 3H), 2.48 (s, 2H), 2.34 (s, 1H),1.66-1.94 (m, 4H), 1.18-1.43 (m, 4H); FXR EC₅₀=958 nM.

Example 1333-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-1-methyl-1H-indazole-6-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 7 with replacement of methyl6-chloro-1-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylate with methyl3-bromo-1-methyl-1H-indazole-6-carboxylate. MS (ESI) m/z: 549.0 [M+H]⁺;¹H NMR (500 MHz, Methanol-d₄) δ 8.19-8.31 (m, 1H), 7.95 (d, J=1.38 Hz,1H), 7.53-7.70 (m, 3H), 7.49 (d, J=9.35 Hz, 1H), 5.97 (s, 1H), 4.53 (s,3H), 3.52-3.85 (m, 5H), 2.61 (s, 2H), 2.36 (s, 1H), 1.97-2.22 (m, 5H),1.09-1.51 (m, 5H); FXR EC₅₀=1175 nM.

Example 1343-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-1-methyl-1H-indazole-6-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 133 with replacement of 2,6-dichlorobenzaldehydewith 2-(trifluoromethyl)benzaldehyde. MS (ESI) m/z: 549.1 [M+H]⁺; ¹H NMR(500 MHz, Methanol-d₄) δ 8.07-8.17 (m, 1H), 7.86-7.96 (m, 1H), 7.72-7.85(m, 3H), 7.69 (br d, J=8.80 Hz, 1H), 7.52 (d, J=6.88 Hz, 1H), 5.74 (s,1H), 3.95 (s, 3H), 3.43-3.71 (m, 4H), 3.12-3.28 (m, 2H), 2.41 (s, 2H),2.31 (s, 1H), 1.58-1.97 (m, 5H), 1.06-1.33 (m, 5H); FXR EC₅₀=3314 nM.

Example 1352-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)pyrrolo[2,1-f][1,2,4]triazine-5-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 123 with replacement of 2,6-dichlorobenzaldehydewith 2-(trifluoromethyl)benzaldehyde. MS (ESI) m/z: 536.1 [M+H]⁺; ¹H NMR(400 MHz, Methanol-d₄) δ 9.00-9.33 (m, 2H), 8.10 (d, J=9.46 Hz, 1H),7.87 (d, J=1.76 Hz, 1H), 7.67-7.83 (m, 3H), 7.51 (dd, J=1.65, 7.15 Hz,1H), 7.14 (d, J=2.20 Hz, 1H), 5.73 (s, 1H), 3.76-4.07 (m, 2H), 3.58 (brdd, J=6.05, 13.75 Hz, 2H), 2.46 (s, 2H), 2.31 (s, 1H), 2.05 (s, 1H),1.76 (t, J=5.61 Hz, 4H), 1.10-1.38 (m, 5H); FXR EC₅₀=87 nM.

Example 1362-(2-(3-(2-Chloro-6-fluorophenyl)-5-cyclopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of 2,6-dichlorobenzaldehydewith 2-chloro-6-fluorobenzaldehyde. MS (ESI) m/z: 554.2 [M+H]⁺; ¹H NMR(500 MHz, DMSO-d₆) δ 8.18 (d, J=1.5 Hz, 1H), 7.68-7.61 (m, 1H), 7.58 (d,J=11.5 Hz, 1H), 7.53 (d, J=8.2 Hz, 1H), 7.42 (t, J=8.7 Hz, 1H), 5.93 (s,1H), 3.54-3.45 (m, 2H), 2.40 (s, 2H), 2.33 (dt, J=3.4, 8.3 Hz, 1H), 1.65(q, J=4.8, 5.3 Hz, 4H), 1.25-1.08 (m, 4H), additional signals missingdue to water signal suppression; FXR EC₅₀=37 nM.

Example 1372-(2-(5-Cyclopropyl-3-(2,6-difluorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of 2,6-dichlorobenzaldehydewith 2,6-difluorobenzaldehyde. MS (ESI) m/z: 538.1 [M+H]⁺; ¹H NMR (500MHz, DMSO-d₆) δ 8.20 (br s, 1H), 7.75-7.50 (m, 2H), 7.33 (t, J=8.1 Hz,2H), 6.02 (s, 1H), 3.79-3.67 (m, 2H), 2.47 (s, 2H), 2.38-2.27 (m, 1H),1.71-1.65 (m, 1H), 1.18 (dtd, J=3.6, 6.5, 29.1 Hz, 4H), additionalsignals missing due to water signal suppression; FXR EC₅₀=155 nM.

Example 1382-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)pyridin-3-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of 2,6-dichlorobenzaldehydewith 2-(trifluoromethyl)nicotinaldehyde. MS (ESI) m/z: 571.3 [M+H]⁺; ¹HNMR (500 MHz, DMSO-d₆) δ 8.92 (d, J=4.8 Hz, 1H), 8.17 (d, J=1.5 Hz, 1H),8.12 (d, J=7.9 Hz, 1H), 7.87 (dd, J=4.8, 7.9 Hz, 1H), 7.57 (dd, J=1.5,11.4 Hz, 1H), 5.83 (s, 1H), 3.69 (br s, 2H), 3.54-3.40 (m, 2H),2.38-2.26 (m, 3H), 1.64-1.57 (m, 4H), 1.28-1.06 (m, 4H); FXR EC₅₀=319nM.

Example 1392-(2-(5-Cyclopropyl-3-(4-(trifluoromethyl)pyridin-3-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 1 with replacement of 2,6-dichlorobenzaldehydewith 4-(trifluoromethyl)nicotinaldehyde. MS (ESI) m/z: 571.1 [M+H]⁺; ¹HNMR (500 MHz, DMSO-d₆) δ 9.01 (d, J=5.2 Hz, 1H), 8.79 (s, 1H), 8.14 (d,J=1.6 Hz, 1H), 7.94 (d, J=5.2 Hz, 1H), 7.61-7.51 (m, 1H), 5.84 (s, 1H),3.76-3.65 (m, 2H), 2.33 (s, 2H), 2.32-2.26 (m, 1H), 1.63 (br s, 4H),1.25-1.08 (m, 4H), additional signals missing due to water signalsuppression; FXR EC₅₀=482 nM.

2-(2-(5-Cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)quinoline-6-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 122 with replacement of 2,6-dichlorobenzaldehydewith 2-(trifluoromethoxy)benzaldehyde. MS (ESI) m/z: 562.1 [M+H]⁺; ¹HNMR (500 MHz, DMSO-d₆) δ 8.30 (s, 1H), 8.09 (d, J=9.34 Hz, 1H),7.88-8.05 (m, 1H), 7.68 (br t, J=7.70 Hz, 1H), 7.49-7.57 (m, 4H), 7.25(br d, J=9.34 Hz, 1H), 5.79-6.01 (m, 1H), 3.95 (br d, J=12.79 Hz, 2H),3.75 (br s, 2H), 3.43-3.64 (m, 1H), 2.40 (s, 2H), 2.21-2.37 (m, 1H),1.57 (br s, 4H), 1.12-1.27 (m, 4H); FXR EC₅₀=133 nM.

Example 1416-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-1-methyl-1H-indole-3-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 8 with replacement of 2,6-dichlorobenzaldehydewith 2-(trifluoromethyl)benzaldehyde. MS (ESI) m/z: 548.1 [M+H]⁺; H NMR(400 MHz, Methanol-d₄) δ 8.27 (d, J=8.58 Hz, 1H), 8.12 (s, 1H),7.89-7.96 (m, 1H), 7.87 (d, J=1.98 Hz, 1H), 7.75-7.82 (m, 2H), 7.51-7.58(m, 1H), 7.46 (dd, J=2.20, 8.80 Hz, 1H), 5.55-6.26 (m, 1H), 3.95 (s,3H), 3.59-3.82 (m, 4H), 2.59 (s, 2H), 2.34 (s, 1H), 2.01-2.21 (m, 5H),1.01-1.45 (m, 4H); FXR EC₅₀=142 nM.

Example 1426-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(2-hydroxyethoxy)quinoline-2-carboxylicacid

Step 1. Methyl 6-bromo-4-(2-hydroxyethoxy)quinoline-2-carboxylate

Methyl 6-bromo-4-hydroxyquinoline-2-carboxylate (0.25 g, 0.87 mmol),2-bromoethan-1-ol (0.19 mL, 2.7 mmol) and potassium carbonate (0.37 g,2.7 mmol) in acetonitrile (15 mL) were heated to 80° C. After 16 hours,additional 2-bromoethan-1-ol (0.19 mL, 2.7 mmol) was added. After 16hours, the reaction mixture was diluted with water (25 mL), extractedwith ethyl acetate (2×25 mL). The organic layer was dried over Na₂SO₄,filtered, and concentrated in vacuo. The residue was purified usingsilica gel chromatography to isolate methyl6-bromo-4-(2-hydroxyethoxy)quinoline-2-carboxylate (0.25 g, 0.78 mmol,88% yield). ¹H NMR (400 MHz, DMSO-d₆) δ 8.51 (d, J=2.2 Hz, 1H),8.06-8.01 (m, 1H), 8.00-7.93 (m, 1H), 7.58 (s, 1H), 4.48 (s, 1H), 4.35(t, J=4.5 Hz, 2H), 3.95 (s, 3H), 3.91-3.84 (m, 2H).

Example 142.6-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(2-hydroxyethoxy)quinoline-2-carboxylicacid

The title compound was prepared as described for the preparation ofExample 85 with replacement of ethyl6-chloro-4-(trifluoromethyl)quinoline-2-carboxylate with methyl6-bromo-4-(2-hydroxyethoxy)quinoline-2-carboxylate. MS (ESI) m/z: 606.2[M+H]⁺; ¹H NMR (500 MHz, Methanol-d₄) δ 8.74 (s, 2H), 8.13 (d, J=9.63Hz, 1H), 7.83-7.93 (m, 1H), 7.73 (s, 1H), 7.56-7.65 (m, 1H), 5.89-5.97(m, 1H), 4.52-4.66 (m, 2H), 4.11 (br d, J=3.85 Hz, 2H), 3.66 (s, 2H),3.41 (s, 2H), 2.50 (s, 2H), 2.30-2.38 (m, 1H), 1.73-1.91 (m, 4H),1.28-1.55 (m, 4H); FXR EC₅₀=147 nM.

Example 1436-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(2-hydroxyethoxy)quinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 142 with replacement of3,5-dichloroisonicotinaldehyde with 2-(trifluoromethyl)benzaldehyde. MS(ESI) m/z: 606.2 [M+H]⁺; ¹H NMR (500 MHz, Methanol-d₄) δ 7.84-7.99 (m,2H), 7.76 (t, J=8.12 Hz, 2H), 7.48-7.58 (m, 4H), 5.65-5.90 (m, 1H), 4.36(t, J=4.54 Hz, 2H), 3.97-4.14 (m, 2H), 3.42-3.48 (m, 2H), 3.12-3.19 (m,2H), 2.39 (s, 2H), 2.24-2.36 (m, 1H), 1.63-1.89 (m, 4H), 1.00-1.33 (m,4H); FXR EC₅₀=159 nM.

Example 1446-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)pyridin-3-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-methoxyquinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 108 with replacement of3,5-dichloroisonicotinaldehyde with 2-(trifluoromethyl)nicotinaldehyde.MS (ESI) m/z: 577.3 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ 8.88 (d, J=4.7Hz, 1H), 8.13-7.96 (m, 2H), 7.85 (dd, J=4.7, 7.9 Hz, 1H), 7.67 (br s,1H), 7.49 (s, 1H), 7.22 (br s, 1H), 5.76 (s, 1H), 4.08 (s, 3H), 3.42 (brs, 2H), 3.13 (br s, 2H), 2.29 (br s, 3H), 1.59 (br s, 4H), 1.23-1.05 (m,4H); FXR EC₅₀=156 nM.

Example 1456-(2-(3-(3-Chloropyridin-2-yl)-5-cyclopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-methoxyquinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 108 with replacement of3,5-dichloroisonicotinaldehyde with 3-chloropicolinaldehyde. MS (ESI)m/z: 543.1 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ 8.69 (d, J=4.6 Hz, 1H),8.15 (d, J=8.2 Hz, 1H), 7.95 (d, J=9.4 Hz, 1H), 7.68 (dd, J=2.8, 9.4 Hz,1H), 7.63 (dd, J=4.7, 8.3 Hz, 1H), 7.47 (s, 1H), 7.26 (d, J=2.7 Hz, 1H),5.87 (s, 1H), 4.09 (s, 3H), 3.18 (br t, J=9.4 Hz, 2H), 2.34 (br s, 3H),1.74-1.59 (m, 4H), 1.24-1.09 (m, 4H), additional signals missing due towater signal suppression; FXR EC₅₀=160 nM.

Example 1466-(2-(5-Cyclopropyl-3-(4-(trifluoromethyl)pyridin-3-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-methoxyquinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 108 with replacement of3,5-dichloroisonicotinaldehyde with 4-(trifluoromethyl)nicotinaldehyde.MS (ESI) m/z: 577.2 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ 9.02 (d, J=5.2Hz, 1H), 8.82 (s, 1H), 8.02-7.87 (m, 2H), 7.63 (dd, J=2.8, 9.4 Hz, 1H),7.46 (s, 1H), 7.27 (d, J=2.8 Hz, 1H), 5.83 (s, 1H), 4.08 (s, 3H), 2.32(br s, 3H), 1.71-1.59 (m, 4H), δ 1.25-1.08 (m, 4H), additional signalsmissing due to water signal suppression; FXR EC₅₀=185 nM.

Example 1476-(2-(3-Cyclohexyl-5-cyclopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-methoxyquinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 108 with replacement of3,5-dichloroisonicotinaldehyde with cyclohexanecarbaldehyde. MS (ESI)m/z: 514.1 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ 7.97 (d, J=9.2 Hz, 1H),7.78-7.68 (m, 1H), 7.49 (s, 1H), 7.33 (d, J=2.8 Hz, 1H), 6.41 (s, 1H),4.12 (s, 3H), 3.58-3.66 (m, 2H), 3.34-3.26 (m, 2H), 2.73 (br s, 3H),2.24-2.17 (m, 1H), 2.00-1.62 (m, 8H), 1.48-1.18 (m, 6H), 1.11-0.95 (m,4H); FXR EC₅₀=1063 nM.

Example 1487-(2-(3-(2,6-Dichlorophenyl)-5-isopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)cinnoline-3-carboxylic acid

The title compound was prepared as described in General Method B for thepreparation of Example 53 with replacement of cyclopropylacetylene withisopropylacetylene. MS (ESI) m/z: 549.1 [M+H]⁺; ¹H NMR (500 MHz, CDCl₃)δ 8.52 (br dd, J=4.8, 1.3 Hz, 1H), 7.85-7.74 (m, 1H), 7.72-7.62 (m, 1H),7.62-7.52 (m, 1H), 7.49-7.37 (m, 2H), 7.37-7.28 (m, 1H), 5.77 (s, 1H),3.69-3.58 (m, 1H), 3.46-3.37 (m, 1H), 3.37-3.25 (m, 2H), 2.59 (s, 2H),2.35 (s, 1H), 1.78 (br s, 4H), 1.43 (br d, J=6.9 Hz, 6H); FXR EC₅₀=548nM.

Example 1496-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)quinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 20 with replacement of 2,6-dichlorobenzaldehydewith 3,5-dichloroisonicotinaldehyde. MS (ESI) m/z: 547.3 [M+H]⁺; ¹H NMR(500 MHz, DMSO-d₆) δ 8.86 (s, 2H), 8.18 (d, J=8.5 Hz, 1H), 7.94 (dd,J=12.7, 9.0 Hz, 2H), 7.75-7.57 (m, 1H), 7.21 (br d, J=1.5 Hz, 1H), 6.00(s, 1H), 3.63-3.46 (m, 2H), 3.00 (s, 1H), 2.40 (s, 2H), 2.39-2.31 (m,1H), 1.79-1.61 (m, 4H), 1.31-1.21 (m, 4H), 1.17 (br d, J=2.6 Hz, 2H);FXR EC₅₀=205 nM.

Example 1506-(2-(3-(3-Chloropyridin-4-yl)-5-cyclopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)quinoline-2-carboxylicacid

The title compound was obtained as a minor isolate during thepreparation of Example 149 from reduction of one chlorine during thePd-catalyzed Buchwald coupling step. MS (ESI) m/z: 513.3 [M+H]⁺; ¹H NMR(500 MHz, DMSO-d₆) δ 8.84 (s, 1H), 8.69 (br d, J=4.9 Hz, 1H), 8.20 (brd, J=8.4 Hz, 1H), 7.94 (br dd, J=12.1, 9.0 Hz, 2H), 7.68 (br d, J=6.9Hz, 1H), 7.60 (d, J=4.8 Hz, 1H), 7.22 (br s, 1H), 7.14 (br s, 2H), 6.01(s, 1H), 3.65-3.44 (m, 3H), 2.46-2.39 (m, 2H), 2.39-2.27 (m, 2H),1.80-1.61 (m, 3H), 1.28-1.17 (m, 2H), 1.19-1.09 (m, 2H); FXR EC₅₀=4345nM.

Example 1516-(2-(5-Cyclopropyl-3-(2,6-difluorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)quinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 20 with replacement of 2,6-dichlorobenzaldehydewith 2,6-difluorobenzaldehyde. MS (ESI) m/z: 514.1 [M+H]⁺; ¹H NMR (500MHz, DMSO-d₆) δ 8.22 (d, J=8.6 Hz, 1H), 7.95 (dd, J=9.0, 13.8 Hz, 2H),7.74-7.63 (m, 2H), 7.33 (t, J=8.1 Hz, 2H), 7.23 (d, J=2.6 Hz, 1H), 6.00(s, 1H), 3.59-3.51 (m, 2H), 3.22 (dd, J=12.1, 22.7 Hz, 2H), 2.46 (s,2H), 2.35 (ddd, J=5.2, 8.5, 13.2 Hz, 1H), 1.80-1.60 (m, 4H), 1.28-1.09(m, 4H); FXR EC₅₀=1991 nM.

Example 1526-(2-(3-(3-Chloropyridin-2-yl)-5-cyclopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)quinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 20 with replacement of 2,6-dichlorobenzaldehydewith 3-chloropicolinaldehyde. MS (ESI) m/z: 513.2 [M+H]⁺; ¹H NMR (500MHz, DMSO-d₆) δ 8.68 (br d, J=4.0 Hz, 1H), 8.21 (br s, 1H), 8.13 (br d,J=8.0 Hz, 1H), 7.95 (br s, 1H), 7.68 (br d, J=5.7 Hz, 1H), 7.62 (dd,J=8.0, 4.6 Hz, 1H), 7.22 (br s, 1H), 5.91 (br s, 1H), 3.51 (br s, 1H),3.26 (br d, J=5.7 Hz, 2H), 2.41-2.25 (m, 3H), 1.69 (br s, 4H), 1.20 (brd, J=7.7 Hz, 2H), 1.14 (br d, J=2.0 Hz, 2H) additional peaks lost underDMSO signal; FXR EC₅₀=1633 nM.

Example 1536-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)pyridin-3-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)quinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 20 with replacement of 2,6-dichlorobenzaldehydewith 2-(trifluoromethyl)nicotinaldehyde. MS (ESI) m/z: 547.3 [M+H]⁺; ¹HNMR (500 MHz, DMSO-d₆) δ 8.89 (d, J=4.8 Hz, 1H), 8.19 (d, J=8.6 Hz, 1H),8.09 (d, J=7.9 Hz, 1H), 7.94 (d, J=8.7 Hz, 2H), 7.85 (dd, J=4.8, 7.9 Hz,1H), 7.65 (d, J=9.4 Hz, 1H), 7.17 (s, 1H), 5.78 (s, 1H), 3.52-3.42 (m,2H), 3.20-3.12 (m, 2H), 2.29 (br s, 3H), 1.67-1.53 (m, 4H), 1.26-1.06(m, 4H); FXR EC₅₀=1282 nM.

Example 1546-(2-(5-Cyclopropyl-3-(4-(trifluoromethyl)pyridin-3-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)quinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 20 with replacement of 2,6-dichlorobenzaldehydewith 4-(trifluoromethyl)nicotinaldehyde. MS (ESI) m/z: 547.3 [M+H]⁺; ¹HNMR (500 MHz, DMSO-d₆) δ 9.03 (d, J=5.1 Hz, 1H), 8.85 (s, 1H), 8.18 (d,J=8.6 Hz, 1H), 7.99-7.88 (m, 3H), 7.66 (dd, J=2.5, 9.4 Hz, 1H), 7.19 (d,J=2.7 Hz, 1H), 5.82 (s, 1H), 3.22-3.16 (m, 2H), 2.36-2.19 (m, 3H),1.70-1.58 (m, 4H), 1.24-1.11 (m, 4H), additional signals missing due towater signal suppression; FXR EC₅₀=554 nM.

Example 1556-(2-(5-Cyclopropyl-3-(2,6-dichloro-4-fluorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)quinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 20 with replacement of 2,6-dichlorobenzaldehydewith 2,6-dichloro-4-fluorobenzaldehyde. MS (ESI) m/z: 564.1 [M+H]⁺; ¹HNMR (500 MHz, DMSO-d₆) δ 8.20 (br d, J=8.5 Hz, 1H), 7.96 (br t, J=9.6Hz, 2H), 7.77 (d, J=8.5 Hz, 2H), 7.69 (br d, J=9.8 Hz, 1H), 7.22 (br s,1H), 5.93 (s, 1H), 3.91 (s, 1H), 3.54 (br d, J=8.2 Hz, 1H), 3.35-3.07(m, 2H), 2.42-2.28 (m, 3H), 1.81-1.61 (m, 4H), 1.22 (br d, J=7.9 Hz,2H), 1.15 (br d, J=2.7 Hz, 2H) additional signals lost due to watersuppression in ¹H NMR experiment; FXR EC₅₀=523 nM.

Example 1566-(2-(3-(2-Chloro-6-fluorophenyl)-5-cyclopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)quinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 20 with replacement of 2,6-dichlorobenzaldehydewith 2-chloro-6-fluorobenzaldehyde. MS (ESI) m/z: 530.4 [M+H]⁺; ¹H NMR(500 MHz, DMSO-d₆) δ 8.20 (d, J=8.6 Hz, 1H), 7.95 (dd, J=9.0, 15.1 Hz,2H), 7.66 (q, J=7.4, 10.2 Hz, 2H), 7.55 (d, J=8.1 Hz, 1H), 7.45 (t,J=8.7 Hz, 1H), 7.22 (s, 1H), 5.92 (s, 1H), 3.57-3.48 (m, 2H), 3.21 (brs, 2H), 2.40 (s, 2H), 2.39-2.28 (m, 1H), 1.76-1.60 (m, 4H), 1.26-1.08(m, 4H); FXR EC₅₀=326 nM.

Example 1576-(2-(3-(2-Chloro-6-methylphenyl)-5-cyclopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)quinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 20 with replacement of 2,6-dichlorobenzaldehydewith 2-chloro-6-methylbenzaldehyde. MS (ESI) m/z: 526.4 [M+H]⁺; ¹H NMR(500 MHz, DMSO-d₆) δ 8.21 (d, J=8.7 Hz, 1H), 8.01-7.88 (m, 2H), 7.68 (d,J=9.4 Hz, 1H), 7.47-7.41 (m, 2H), 7.35 (t, J=4.5 Hz, 1H), 7.21 (s, 1H),5.76 (s, 1H), 3.24-3.14 (m, 2H), 2.35 (q, J=12.5 Hz, 3H), 2.10 (s, 3H),1.72-1.58 (m, 4H), 1.25-1.08 (m, 4H), additional signals missing due towater signal suppression; FXR EC₅₀=223 nM.

Example 1587-(2-(3-(2-Chloro-6-fluorophenyl)-5-cyclopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)isoquinoline-3-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 129 with replacement of 2,6-dichlorobenzaldehydewith 2-chloro-6-fluorobenzaldehyde. MS (ESI) m/z: 530.2 [M+H]⁺; ¹H NMR(500 MHz, DMSO-d₆) δ 9.09 (s, 1H), 8.40 (s, 1H), 7.96 (d, J=9.1 Hz, 1H),7.73-7.59 (m, 2H), 7.54 (d, J=8.2 Hz, 1H), 7.50-7.36 (m, 2H), 5.91 (s,1H), 3.28-3.20 (m, 2H), 2.40 (s, 2H), 2.34 (tt, J=5.0, 8.2 Hz, 1H),1.73-1.59 (m, 4H), 1.27-1.09 (m, 4H), additional signals missing due towater signal suppression; FXR EC₅₀=459 nM.

Example 1597-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)isoquinoline-3-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 129 with replacement of 2,6-dichlorobenzaldehydewith 2-(trifluoromethyl)benzaldehyde. MS (ESI) m/z: 546.1 [M+H]⁺; ¹H NMR(500 MHz, DMSO-d₆) δ 9.09 (s, 1H), 8.39 (s, 1H), 7.93 (dd, J=13.4, 8.4Hz, 2H), 7.85-7.73 (m, 2H), 7.66 (dd, J=9.2, 2.3 Hz, 1H), 7.56 (d, J=7.3Hz, 1H), 7.39 (d, J=1.7 Hz, 1H), 5.78 (s, 1H), 3.52 (br dd, J=12.6, 5.2Hz, 2H), 2.36-2.20 (m, 3H), 1.75-1.56 (m, 4H), 1.26-1.17 (m, 2H),1.16-1.08 (m, 2H), 1.02 (d, J=6.2 Hz, 1H) additional proton signals werelost due to water suppression; FXR EC₅₀=254 nM.

Example 1607-(2-(5-Cyclopropyl-3-(4-(trifluoromethyl)pyridin-3-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)isoquinoline-3-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 129 with replacement of 2,6-dichlorobenzaldehydewith 4-(trifluoromethyl)nicotinaldehyde. MS (ESI) m/z: 547.2 [M+H]⁺; ¹HNMR (500 MHz, DMSO-d₆) δ 9.08 (s, 1H), 9.03 (d, J=5.2 Hz, 1H), 8.85 (s,1H), 8.38 (s, 1H), 7.97 (d, J=5.2 Hz, 1H), 7.94 (d, J=9.2 Hz, 1H),7.70-7.64 (m, 1H), 7.39 (s, 1H), 5.82 (s, 1H), 3.26-3.18 (m, 2H),2.37-2.29 (m, 3H), 1.72-1.57 (m, 4H), 1.25-1.10 (m, 4H), additionalsignals missing due to water signal suppression; FXR EC₅₀=1608 nM.

Example 1617-(2-(3-(3-Chloropyridin-2-yl)-5-cyclopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)isoquinoline-3-carboxylic acid

The title compound was prepared as described in General Method B for thepreparation of Example 129 with replacement of 2,6-dichlorobenzaldehydewith 4-(trifluoromethyl)nicotinaldehyde. MS (ESI) m/z: 513.2 [M+H]⁺; ¹HNMR (500 MHz, DMSO-d₆) δ 9.10 (s, 1H), 8.69 (br d, J=4.6 Hz, 1H), 8.40(s, 1H), 8.15 (br d, J=7.9 Hz, 1H), 7.96 (br d, J=9.2 Hz, 1H), 7.69 (brd, J=8.2 Hz, 1H), 7.63 (dd, J=8.2, 4.6 Hz, 1H), 7.41 (s, 1H), 5.88 (s,1H), 3.67-3.41 (m, 1H), 3.37-3.12 (m, 2H), 2.41-2.22 (m, 3H), 1.81-1.55(m, 5H), 1.21 (br d, J=7.9 Hz, 2H), 1.14 (br d, J=2.4 Hz, 2H); FXREC₅₀=2324 nM.

Example 1627-(2-(5-Cyclopropyl-3-(2,6-difluorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)isoquinoline-3-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 129 with replacement of 2,6-dichlorobenzaldehydewith 2,6-difluorobenzaldehyde. MS (ESI) m/z: 514.1 [M+H]⁺; ¹H NMR (500MHz, DMSO-d₆) δ 9.10 (s, 1H), 8.41 (s, 1H), 7.96 (d, J=9.1 Hz, 1H), 7.70(q, J=6.7, 7.3 Hz, 2H), 7.42 (s, 1H), 7.33 (t, J=8.2 Hz, 2H), 6.00 (s,1H), 3.61-3.53 (m, 2H), 3.29-3.21 (m, 2H), 2.46 (s, 2H), 2.39-2.28 (m,1H), 1.77-1.57 (m, 4H), 1.28-1.08 (m, 4H); FXR EC₅₀=2824 nM.

Example 1636-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)nicotinicacid

The title compound was prepared as described in General Method A for thepreparation of Example 116 with replacement of 2,6-dichlorobenzaldehydewith 2-(trifluoromethyl)benzaldehyde. MS (ESI) m/z: 564.4 [M+H]⁺; ¹H NMR(500 MHz, DMSO-d₆) δ 8.63 (s, 1H), 7.92 (br d, J=7.93 Hz, 1H), 7.79 (brdd, J=7.63, 11.90 Hz, 2H), 7.56 (br d, J=7.02 Hz, 1H), 7.02 (s, 1H),5.75 (s, 1H), 3.72-4.11 (m, 2H), 2.55 (s, 2H), 2.29 (s, 1H), 1.41-1.64(m, 4H), 1.15-1.24 (m, 2H), 1.11 (br d, J=2.44 Hz, 2H), additionalsignals missing due to water signal suppression; FXR EC₅₀=344 nM.

Example 1646-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-1-methyl-1H-indazole-3-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 125 with replacement of 2,6-dichlorobenzaldehydewith 2-(trifluoromethyl)benzaldehyde. MS (ESI) m/z: 549.1 [M+H]⁺; ¹H NMR(400 MHz, Methanol-d₄) δ 8.27 (d, J=8.58 Hz, 1H), 8.12 (s, 1H),7.89-7.96 (m, 1H), 7.87 (d, J=1.98 Hz, 1H), 7.75-7.82 (m, 2H), 7.51-7.58(m, 1H), 7.46 (dd, J=2.20, 8.80 Hz, 1H), 5.55-6.26 (m, 1H), 3.95 (s,3H), 3.59-3.82 (m, 4H), 2.59 (s, 2H), 2.34 (s, 1H), 2.01-2.21 (m, 5H),1.01-1.45 (m, 4H); FXR EC₅₀=392 nM.

Example 1656-(2-(5-Cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-1-methyl-1H-indazole-3-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 125 with replacement of 2,6-dichlorobenzaldehydewith 2-(trifluoromethoxy)benzaldehyde. MS (ESI) m/z: 565.3 [M+H]⁺; ¹HNMR (500 MHz, DMSO-d₆) δ 7.74-7.67 (m, 1H), 7.65 (br d, J=9.2 Hz, 1H),7.62-7.47 (m, 4H), 7.38 (br d, J=8.5 Hz, 1H), 5.95 (s, 1H), 4.09 (s,3H), 3.27 (br d, J=4.6 Hz, 1H), 3.03 (br s, 1H), 2.41 (s, 2H), 2.37-2.27(m, 1H), 1.82-1.64 (m, 4H), 1.24 (s, 2H), 1.21-1.15 (m, 2H), 1.12 (br d,J=2.4 Hz, 2H); FXR EC₅₀=2975 nM.

Example 1666-(2-(5-Cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-3-(trifluoromethyl)imidazo[1,5-a]pyridine-1-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 126 with replacement of 2,6-dichlorobenzaldehydewith 2-(trifluoromethoxy)benzaldehyde. MS (ESI) m/z: 619.1 [M+H]⁺; ¹HNMR (500 MHz, DMSO-d₆) δ 8.04 (d, J=9.77 Hz, 1H), 7.70 (s, 1H),7.47-7.61 (m, 5H), 5.82-5.99 (m, 1H), 3.20-3.50 (m, 2H), 3.00 (br s,2H), 2.39 (s, 2H), 2.23-2.35 (m, 1H), 1.56-1.86 (m, 4H), 1.05-1.33 (m,4H); FXR EC₅₀=494 nM.

Example 1676-(2-(3-Cyclohexyl-5-cyclopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-ethoxyquinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 72 with replacement of2-(trifluoromethyl)benzaldehyde with cyclohexanecarbaldehyde. MS (ESI)m/z: 528.4 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ 7.99 (d, J=9.3 Hz, 1H),7.76 (d, J=9.2 Hz, 1H), 7.48 (s, 1H), 7.34 (d, J=2.8 Hz, 1H), 6.41 (s,1H), 4.41 (q, J=6.9 Hz, 2H), 3.66-3.55 (m, 2H), 3.33-3.23 (m, 2H), 2.73(br s, 3H), 2.24-2.16 (m, 1H), 1.17-1.61 (m, 8H), 1.51 (t, J=6.9 Hz,3H), 1.47-1.16 (m, 6H), 1.11-0.94 (m, 4H); FXR EC₅₀=3096 nM.

Example 1686-(2-(3-Cyclohexyl-5-cyclopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-1-methyl-1H-indole-3-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 8 with replacement of 2,6-dichlorobenzaldehydewith cyclohexanecarbaldehyde. MS (ESI) m/z: 486.4 [M+H]⁺; ¹H NMR (500MHz, DMSO-d₆) δ 7.94 (d, J=7.4 Hz, 2H), 7.31 br (s, 1H), 7.19 (br s,1H), 6.43 (s, 1H), 3.83 (s, 3H), 3.50 (br s, 2H), 2.77-2.69 (m, 3H),2.20 (tt, J=5.1, 8.7 Hz, 1H), 2.04-1.64 (m, 8H), 1.52-1.19 (m, 6H),1.12-0.94 (m, 4H), additional signals missing due to water signalsuppression; FXR EC₅₀=3972 nM.

Example 1692-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-6-methylpyrimidine-4-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 120 with replacement of 2,6-dichlorobenzaldehydewith 2-(trifluoromethyl)benzaldehyde. MS (ESI) m/z: 511.3 [M+H]⁺; ¹H NMR(500 MHz, DMSO-d₆) δ 7.92 (br d, J=7.63 Hz, 1H), 7.70-7.87 (m, 2H), 7.56(br d, J=7.32 Hz, 1H), 6.82 (s, 1H), 5.64-5.92 (m, 1H), 3.97 (br d,J=13.12 Hz, 2H), 2.55 (m, 2H), 2.28 (s, 3H), 1.91 (s, 1H), 1.47 (br s,4H), 1.15-1.26 (m, 2H), 1.11 (br d, J=2.14 Hz, 2H), additional signalsmissing due to water signal suppression; FXR EC₅₀=1300 nM.

Example 1707-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-8-methylquinoline-3-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 132 with replacement of 2,6-dichlorobenzaldehydewith 2-(trifluoromethyl)benzaldehyde. MS (ESI) m/z: 560.0 [M+H]⁺; ¹H NMR(500 MHz, Methanol-d₄) δ 8.10-8.23 (m, 1H), 8.04 (d, J=8.53 Hz, 1H),7.93 (d, J=9.35 Hz, 1H), 7.44-7.67 (m, 3H), 7.18 (d, J=2.20 Hz, 1H),5.85 (s, 1H), 4.87 (s, 3H), 3.48 (br dd, J=5.09, 11.97 Hz, 2H), 3.33 (brs, 2H), 3.04-3.25 (m, 2H), 2.42 (s, 2H), 2.18-2.35 (m, 1H), 1.61-1.97(m, 4H), 1.04-1.36 (m, 4H); FXR EC₅₀=1747 nM.

Example 1716-(2-(3-(3-Chloropyridin-2-yl)-5-cyclopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)nicotinicacid

The title compound was prepared as described in General Method A for thepreparation of Example 2 with replacement of 2,6-dichlorobenzaldehydewith 3-chloropicolinaldehyde. MS (ESI) m/z: 463.2 [M+H]⁺; ¹H NMR (500MHz, DMSO-d₆) δ 8.68 (d, J=4.6 Hz, 1H), 8.58 (d, J=1.5 Hz, 1H), 8.13 (d,J=8.2 Hz, 1H), 7.89 (dd, J=9.2, 1.8 Hz, 1H), 7.62 (dd, J=8.2, 4.9 Hz,1H), 6.82 (d, J=9.2 Hz, 1H), 5.86 (s, 1H), 3.88-3.76 (m, 1H), 3.45-3.33(m, 1H), 3.27 (dd, J=10.5, 6.0 Hz, 1H), 3.21-3.11 (m, 1H), 2.41-2.24 (m,3H), 1.53 (br t, J=5.2 Hz, 4H), 1.29-1.16 (m, 2H), 1.16-1.08 (m, 2H),1.01 (d, J=6.1 Hz, 1H); FXR EC₅₀=1871 nM.

Example 1725-(2-(5-Cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-1-methyl-1H-pyrrolo[2,3-c]pyridine-2-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 17 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with ethyl5-chloro-1-methyl-1H-pyrrolo[2,3-c]pyridine-2-carboxylate. MS (ESI) m/z:565.9 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ 8.46 (s, 1H), 7.66-7.77 (m,1H), 7.47-7.64 (m, 3H), 6.78 (s, 1H), 6.62 (s, 1H), 5.91 (s, 1H), 4.02(s, 3H), 3.52-3.68 (m, 2H), 3.15 (br d, J=9.16 Hz, 2H), 2.38 (m, 3H),1.52-1.78 (m, 4H), 1.09-1.37 (m, 4H); FXR EC₅₀=2366 nM.

Example 1733-(2-(5-Cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-1-methyl-1H-pyrazolo[4,3-b]pyridine-6-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 113 with replacement of methyl6-bromo-4-ethoxyquinoline-2-carboxylate with methyl3-bromo-1-methyl-1H-pyrazolo[4,3-b]pyridine-6-carboxylate. MS (ESI) m/z:566.2 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ 8.78 (s, 1H), 8.40 (s, 1H),7.65-7.74 (m, 2H), 7.40-7.63 (m, 3H), 5.91 (s, 1H), 3.83-3.84 (m, 2H),3.88 (s, 3H), 2.36-2.44 (m, 2H), 2.32 (br s, 1H), 1.59-1.77 (m, 4H),1.15-1.23 (m, 2H), 1.11 (br d, J=2.44 Hz, 2H), additional signalsmissing due to water signal suppression; FXR EC₅₀=2791 nM.

Example 1745-(2-(5-Cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-1-methyl-1H-indazole-3-carboxylicacid

The title compound was obtained via hydrolysis of5-(2-(5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-1-methyl-1H-indazole-3-carbonitrileunder the conditions described in General Method D for the preparationof Example 100. MS (ESI) m/z: 565.1 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ7.67-7.76 (m, 1H), 7.65 (br d, J=9.16 Hz, 1H), 7.48-7.61 (m, 3H), 7.38(br d, J=8.54 Hz, 1H), 7.02-7.31 (m, 1H), 5.95 (s, 1H), 3.27 (br d,J=4.58 Hz, 2H), 3.03 (br s, 2H), 2.55 (s, 3H), 2.37-2.45 (m, 2H), 2.33(br s, 1H), 1.59-1.89 (m, 4H), 1.05-1.21 (m, 4H); FXR EC₅₀=236 nM.

Example 1756-(2-(5-Cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-1-methyl-1H-indazole-3-carbonitrile

A slurry of5-cyclopropyl-4-(7-azaspiro[3.5]non-1-en-2-yl)-3-(2-(trifluoromethoxy)phenyl)isoxazole(18 mg, 0.046 mmol), 6-bromo-1-methyl-1H-indazole-3-carbonitrile (16.3mg, 0.069 mmol) and Cs₂CO₃ (30.0 mg, 0.092 mmol) in dioxane (154 L) wasdegassed by bubbling nitrogen through the mixture for 5 min.Chloro(2-dicyclohexylphosphino-2′,6′-diisopropoxy-1,1′-biphenyl)[2-(2′-amino-1,1′-biphenyl)]palladium(II)(RuPhos-Pd-G2) (1.791 mg, 2.305 μmol) was added and the reaction mixturewas sealed and heated to 90° C. After 24 h, 2 mL of methanol was added,the solids were filtered, and the filtrate was purified via preparativeLC/MS with the following conditions: Column: XBridge C18, 19×200 mm,5-μm particles; Mobile Phase A: 5:95 acetonitrile: water with 0.1%trifluoroacetic acid; Mobile Phase B: 95:5 acetonitrile: water with 0.1%trifluoroacetic acid; Gradient: 35-75% B over 25 minutes, then a5-minute hold at 100% B; Flow: 20 mL/min. Fractions containing thedesired product were combined and dried to give6-(2-(5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-1-methyl-1H-indazole-3-carbonitrile(11.1 mg, 0.019 mmol, 42% yield). MS (ESI) m/z: 545.9 [M+H]⁺; ¹H NMR(500 MHz, DMSO-d₆) δ 7.63-7.80 (m, 4H), 7.48-7.62 (m, 3H), 7.32 (br d,J=9.77 Hz, 1H), 6.85 (s, 1H), 5.80-6.18 (m, 1H), 4.23 (s, 3H), 3.19-3.51(m, 2H), 3.04 (br t, J=9.16 Hz, 2H), 2.35-2.42 (m, 2H), 2.31 (br s, 1H),1.54-1.81 (m, 4H), 1.03-1.33 (m, 4H); FXR EC₅₀=3244 nM.

Example 1766-(2-(5-Cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-1-methyl-1H-indazole-3-carboxylicacid

The title compound was obtained via hydrolysis of6-(2-(5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-1-methyl-1H-indazole-3-carbonitrile(Example 175) under the conditions described in General Method D for thepreparation of Example 100. MS (ESI) m/z: 565.3 [M+H]⁺; ¹H NMR (500 MHz,DMSO-d₆) δ 7.82 (br d, J=8.85 Hz, 1H), 7.64-7.75 (m, 1H), 7.49-7.62 (m,4H), 7.08 (br d, J=8.54 Hz, 1H), 6.94 (br s, 1H), 5.80-6.02 (m, 1H),4.01 (br s, 3H), 3.08 (br t, J=9.16 Hz, 2H), 2.39 (s, 2H), 2.22-2.35 (m,1H), 1.59-1.74 (m, 4H), 1.18 (br d, J=7.93 Hz, 2H), 1.11 (br d, J=2.14Hz, 2H), additional signals missing due to water signal suppression; FXREC₅₀=2975 nM.

Example 1772-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)thiazolo[4,5-b]pyridine-6-carboxylic acid

Step 1.4-(7-(6-Bromothiazolo[4,5-b]pyridin-2-yl)-7-azaspiro[3.5]non-1-en-2-yl)-5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazole

Cesium carbonate (83 mg, 0.26 mmol) was added to a room temp solution of5-cyclopropyl-3-(2,6-dichlorophenyl)-4-(7-azaspiro[3.5]non-1-en-2-yl)isoxazole(50 mg, 0.102 mmol, synthesis described in General Method A) and6-bromo-2-chlorothiazolo[4,5-b]pyridine (38.2 mg, 0.15 mmol) in DMA(0.25 mL). The reaction mixture was heated to 50° C. for 4 h and thecrude reaction mixture was purified by flash chromatography on SiO₂(0-100% EtOAc/hexanes, Isco 12 g column) to give4-(7-(6-bromothiazolo[4,5-b]pyridin-2-yl)-7-azaspiro[3.5]non-1-en-2-yl)-5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazole(51 mg, 0.082 mmol, 81% yield). ¹H NMR (500 MHz, CDCl₃) δ 8.40 (d,J=2.20 Hz, 1H), 7.93 (d, J=1.93 Hz, 1H), 7.41-7.50 (m, 2H), 7.33-7.39(m, 1H), 5.78 (s, 1H), 3.68-3.94 (m, 2H), 3.40-3.63 (m, 2H), 2.42 (s,2H), 2.14-2.28 (m, 1H), 1.67-1.85 (m, 5H), 1.28-1.39 (m, 3H), 1.10-1.22(m, 2H).

Step 2.2-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)thiazolo[4,5-b]pyridine-6-carbonitrile

A microwave vial containing4-(7-(6-bromothiazolo[4,5-b]pyridin-2-yl)-7-azaspiro[3.5]non-1-en-2-yl)-5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazole(30 mg, 0.051 mmol), Xantphos (5.9 mg, 10.2 μmol), Pd₂(dba)₃ (9.3 mg,10.2 μmol), and zinc cyanide (6.0 mg, 0.051 mmol) was purged three timeswith nitrogen and then anhydrous DMF (0.5 mL) was added. The reactionmixture was heated under microwave irradiation at 110° C. for 1.5 h. Thereaction mixture was diluted with EtOAc and washed with brine. Theorganic layer was dried over Na₂SO₄, filtered and concentrated in vacuoto dryness. The residue was purified by flash chromatography on SiO₂(0-100% EtOAc/hexanes, Isco 12 g column) to yield2-(2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)thiazolo[4,5-b]pyridine-6-carbonitrile(15.5 mg, 0.028 mmol, 54% yield) as a gum. ¹H NMR (400 MHz, CDCl₃) δ8.54-8.79 (m, 1H), 7.87-8.19 (m, 1H), 7.42-7.49 (m, 2H), 7.29-7.41 (m,2H), 5.71-5.87 (m, 1H), 3.75-4.02 (m, 2H), 3.63 (br d, J=3.96 Hz, 2H),2.44 (s, 2H), 2.12-2.36 (m, 2H), 1.79 (t, J=5.72 Hz, 4H), 1.62 (br s,3H), 1.29-1.38 (m, 2H), 1.09-1.25 (m, 3H), 0.88 (dd, J=3.30, 7.92 Hz,1H).

Example 177.2-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)thiazolo[4,5-b]pyridine-6-carboxylicacid

The title compound was obtained via hydrolysis of2-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)thiazolo[4,5-b]pyridine-6-carbonitrileunder the conditions described in General Method D for the preparationof Example 100. MS (ESI) m/z: 552.9 [M+H]⁺; ¹H NMR (400 MHz,Methanol-d₄) δ 8.93 (d, J=1.76 Hz, 1H), 8.81 (d, J=1.76 Hz, 1H),7.45-7.70 (m, 4H), 5.87 (s, 1H), 3.58-4.43 (m, 5H), 2.52 (s, 2H), 2.33(s, 1H), 1.83 (t, J=5.72 Hz, 4H), 1.10-1.43 (m, 5H); FXR EC₅₀=121 nM.

6-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)quinoxaline-2-carboxylicacid

Step 1.6-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)quinoxaline-2-carbonitrile

A slurry of5-cyclopropyl-4-(7-azaspiro[3.5]non-1-en-2-yl)-3-(2-(trifluoromethyl)phenyl)isoxazole(30 mg, 0.08 mmol, synthesis described in General method A),6-chloroquinoxaline-2-carbonitrile (18.7 mg, 0.10 mmol) and Cs₂CO₃ (52.2mg, 0.16 mmol) in dioxane (0.40 mL) was degassed by bubbling nitrogenthrough the mixture for 5 min.Chloro(2-dicyclohexylphosphino-2′,6′-diisopropoxy-1,1′-biphenyl)[2-(2′-amino-1,1′-biphenyl)]palladium(II)(RuPhos-Pd-G2) (3.11 mg, 4.01 μmol) was then added and the reactionmixture was sealed and heated to 90° C. for 6 h. The crude mixturepurified directly by flash chromatography on SiO₂ (0-100% EtOAc/hexanes,Isco 12 g column) to yield6-(2-(5-cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)quinoxaline-2-carbonitrile(31 mg, 0.056 mmol, 70% yield) as a gum. MS (ESI) m/z: 528.1 [M+H]⁺; ¹HNMR (400 MHz, CDCl₃) δ 8.83 (s, 1H), 7.89 (d, J=9.7 Hz, 1H), 7.84-7.77(m, 1H), 7.68-7.60 (m, 2H), 7.58 (dd, J=9.6, 2.8 Hz, 1H), 7.44 (dd,J=7.0, 1.8 Hz, 1H), 7.18 (d, J=2.9 Hz, 1H), 5.63 (s, 1H), 3.61 (dt,J=13.4, 5.0 Hz, 2H), 3.38 (ddd, J=13.0, 8.1, 4.4 Hz, 2H), 2.39 (s, 2H),2.15 (tt, J=8.4, 5.1 Hz, 1H), 1.80-1.69 (m, 4H), 1.32-1.26 (m, 2H),1.19-1.12 (m, 2H).

Example 178.6-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)quinoxaline-2-carboxylicacid

The title compound was obtained via hydrolysis of6-(2-(5-cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)quinoxaline-2-carbonitrileunder the conditions described in General Method D for the preparationof Example 100. MS (ESI) m/z: 547.1 [M+H]⁺; ¹H NMR (500 MHz, CDCl₃) δ9.45 (s, 1H), 7.93 (d, J=9.35 Hz, 1H), 7.82 (br d, J=7.43 Hz, 1H),7.55-7.74 (m, 3H), 7.39-7.55 (m, 1H), 7.31 (d, J=1.93 Hz, 1H), 5.65 (s,1H), 3.51-3.66 (m, 2H), 3.31-3.51 (m, 2H), 2.41 (s, 2H), 1.98-2.20 (m,1H), 1.62-1.81 (m, 4H), 1.24-1.37 (m, 2H), 1.09-1.24 (m, 2H); FXREC₅₀=172 nM.

General Method E Example 1792-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-N-(cyclopropylsulfonyl)-4-fluorobenzo[d]thiazole-6-carboxamide

2-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid (Example 1, 15 mg, 0.03 mmol, synthesis described in General MethodA) was dissolved in THF (0.26 mL) in a 5 mL round bottom flask that wasequipped with a magnetic stirrer under nitrogen. CDI (12.8 mg, 0.08mmol) was added and the mixture was heated at 60° C. for 1 h followed byaddition of cyclopropanesulfonamide (12.7 mg, 0.10 mmol) and DBU (11.9μL, 0.08 mmol). The reaction mixture was stirred at room temperature for6 h. The crude mixture was purified via preparative LC/MS with thefollowing conditions: Column: XBridge C18, 19×200 mm, 5-μm particles;Mobile Phase A: 5:95 acetonitrile: water with 0.1% TFA; Mobile Phase B:95:5 acetonitrile: water with 0.1% TFA; Gradient: 5-100% B over 20minutes, then a hold at 100% B; Flow: 20 mL/min. Fractions containingthe desired product were combined and dried to give2-(2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-N-(cyclopropylsulfonyl)-4-fluorobenzo[d]thiazole-6-carboxamide(12.0 mg, 0.02 mmol, 64% yield). MS (ESI) m/z: 673.0 [M+H]⁺; ¹H NMR (400MHz, CDCl₃) δ 8.11 (d, J=1.54 Hz, 1H), 7.65 (dd, J=1.43, 11.99 Hz, 1H),7.36 (d, J=1.76 Hz, 1H), 7.34 (d, J=0.66 Hz, 1H), 7.30 (s, 1H), 5.69 (s,1H), 3.64-3.74 (m, 2H), 3.27-3.35 (m, 2H), 2.06-2.18 (s, 2H), 1.88-1.93(m, 2H), 1.04-1.16 (m, 4H), 0.64-0.92 (m, 8H); FXR EC₅₀=13 nM.

Example 1802-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluoro-N-(methylsulfonyl)benzo[d]thiazole-6-carboxamide

The title compound was prepared as described in General Method E for thepreparation of Example 179 with replacement of cyclopropanesulfonamidewith methanesulfonamide. MS (ESI) m/z: 646.9 [M+H]⁺; ¹H NMR (500 MHz,DMSO-d₆) δ 8.11 (s, 1H), 7.67 (s, 2H), 7.54-7.65 (m, 2H), 6.94-7.35 (m,1H), 5.91 (s, 1H), 2.90 (s, 2H), 2.55 (s, 3H), 2.28-2.44 (s, 2H), 1.92(m, 1H), 1.60-1.71 (m, 4H), 1.22 (br d, J=7.93 Hz, 2H), 1.12-1.18 (m,2H), additional signals missing due to water signal suppression; FXREC₅₀=35 nM.

Example 1812-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-N-(cyclopropylsulfonyl)-4-fluorobenzo[d]thiazole-6-carboxamide

The title compound was prepared as described in General Method E for thepreparation of Example 179 with replacement of 2,6-dichlorobenzaldehydewith 3,5-dichloroisonicotinaldehyde. MS (ESI) m/z: 674.0 [M+H]⁺; ¹H NMR(500 MHz, CDCl₃) δ 8.68 (br s, 1H), 8.66 (s, 2H), 7.92 (d, J=1.38 Hz,1H), 7.53 (dd, J=1.24, 10.87 Hz, 1H), 5.84 (s, 1H), 3.71-3.94 (m, 2H),3.58 (ddd, J=4.54, 7.84, 12.93 Hz, 2H), 2.47 (s, 2H), 2.07-2.30 (m, 2H),1.73-1.88 (m, 4H), 1.47 (dd, J=1.93, 4.68 Hz, 2H), 1.34 (dd, J=2.34,4.81 Hz, 2H), 1.10-1.29 (m, 4H); FXR EC₅₀=60 nM.

2-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluoro-N-(methylsulfonyl)benzo[d]thiazole-6-carboxamide

The title compound was prepared as described in General Method E for thepreparation of Example 181 with replacement of cyclopropanesulfonamidewith methanesulfonamide. MS (ESI) m/z: 648.0 [M+H]⁺; ¹H NMR (400 MHz,CDCl₃) δ 8.65 (s, 2H), 8.03 (d, J=9.24 Hz, 1H), 7.61 (s, 2H), 7.40 (d,J=2.64 Hz, 1H), 5.86 (s, 1H), 4.16 (s, 3H), 3.44-3.59 (m, 2H), 3.12-3.37(m, 2H), 2.45 (s, 2H), 1.82 (br d, J=2.42 Hz, 7H), 1.13-1.44 (m, 8H),0.91 (s, 3H); FXR EC₅₀=688 nM.

Example 1836-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-N-(cyclopropylsulfonyl)-4-(trifluoromethyl)quinoline-2-carboxamide

The title compound was prepared as described in General Method E for thepreparation of Example 179 with replacement of2-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid (Example 1) with6-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylicacid (Example 85). MS (ESI) m/z: 718.2 [M+H]⁺; ¹H NMR (400 MHz,Acetone-d₆) δ 8.62-8.83 (m, 1H), 8.43 (br d, J=1.76 Hz, 1H), 8.11 (brdd, J=1.87, 8.69 Hz, 1H), 7.88 (s, 2H), 7.32-7.69 (m, 2H), 3.89-4.29 (m,2H), 3.47-3.80 (m, 2H), 2.38-2.62 (m, 2H), 2.13-2.28 (m, 1H), 1.65-1.78(m, 1H), 0.95-1.34 (m, 5H); FXR EC₅₀=55 nM.

6-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-N-(methylsulfonyl)-4-(trifluoromethyl)quinoline-2-carboxamide

The title compound was prepared as described in General Method E for thepreparation of Example 183 with replacement of cyclopropanesulfonamidewith methanesulfonamide. MS (ESI) m/z: 691.1 [M+H]⁺; ¹H NMR (400 MHz,Acetone-d₆) δ 8.62-8.83 (m, 1H), 8.43 (br d, J=1.76 Hz, 1H), 8.11 (brdd, J=1.87, 8.69 Hz, 1H), 7.88 (s, 2H), 7.32-7.69 (m, 2H), 3.89-4.29 (m,2H), 3.47-3.80 (m, 2H), 2.38-2.62 (m, 2H), 2.13-2.28 (m, 1H), 1.65-1.78(m, 1H), 0.95-1.34 (m, 5H); FXR EC₅₀=73 nM; Mouse in vivo (3 mg/kg, @ 6h): Cyp7a1=−99%, Fgf15=+28×.

Example 1856-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(difluoromethoxy)-N-(methylsulfonyl)quinoline-2-carboxamide

The title compound was prepared as described in General Method E for thepreparation of Example 184 with replacement of ethyl6-chloro-4-(trifluoromethyl)quinoline-2-carboxylate with methyl6-bromo-4-(difluoromethoxy)quinoline-2-carboxylate. MS (ESI) m/z: 690.0[M+H]⁺; ¹H NMR (500 MHz, Methanol-d₄) δ 8.75 (br s, 2H), 8.14 (br d,J=8.53 Hz, 1H), 7.83 (br s, 2H), 7.10-7.65 (m, 2H), 5.81-6.14 (m, 1H),3.67 (br d, J=5.50 Hz, 2H), 3.42 (br s, 2H), 2.68 (br s, 2H), 2.54 (brs, 3H), 2.23-2.48 (m, 1H), 1.70-1.93 (m, 4H), 1.11-1.56 (m, 4H); FXREC₅₀=16 nM.

6-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(difluoromethoxy)-N-(methylsulfonyl)quinoline-2-carboxamide

The title compound was prepared as described in General Method E for thepreparation of Example 185 with replacement of3,5-dichloroisonicotinaldehyde with 2-(trifluoromethyl)benzaldehyde. MS(ESI) m/z: 689.0 [M+H]⁺; ¹H NMR (500 MHz, Methanol-d₄) δ 8.03 (d, J=9.35Hz, 1H), 7.88 (s, 1H), 7.67-7.81 (m, 4H), 7.51 (d, J=7.15 Hz, 1H),7.20-7.49 (m, 1H), 7.32 (d, J=2.75 Hz, 1H), 5.73 (s, 1H), 3.50-3.68 (m,1H), 3.23-3.32 (m, 2H), 2.42 (s, 2H), 2.31 (s, 1H), 2.05 (s, 3H),1.67-1.87 (m, 4H), 1.14-1.29 (m, 4H); FXR EC₅₀=13 nM.

Example 1876-(2-(5-Cyclopropyl-3-(4-(trifluoromethyl)pyridin-3-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(difluoromethoxy)-N-(methylsulfonyl)quinoline-2-carboxamide

The title compound was prepared as described in General Method E for thepreparation of Example 185 with replacement of3,5-dichloroisonicotinaldehyde with 4-(trifluoromethyl)nicotinaldehyde.MS (ESI) m/z: 690.3 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ 8.93 (d, J=4.7Hz, 1H), 8.15 (d, J=7.9 Hz, 1H), 8.02 (d, J=9.4 Hz, 1H), 7.89 (dd,J=4.8, 7.9 Hz, 1H), 7.86-7.55 (m, 3H), 7.15 (d, J=2.7 Hz, 1H), 3.35 (s,3H), 3.29-3.21 (m, 2H), 2.38-2.28 (m, 3H). 1.66 (q, J=7.1, 7.7 Hz, 4H),1.25-1.08 (m, 4H), additional signals missing due to water signalsuppression; FXR EC₅₀=63 nM.

6-(2-(5-Cyclopropyl-3-(4-(trifluoromethyl)pyridin-3-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-N-(cyclopropylsulfonyl)-4-(difluoromethoxy)quinoline-2-carboxamide

The title compound was prepared as described in General Method E for thepreparation of Example 187 with replacement of methanesulfonamide withcyclopropanesulfonamide. MS (ESI) m/z: 716.3 [M+H]⁺; ¹H NMR (500 MHz,DMSO-d₆) δ 8.92 (d, J=4.6 Hz, 1H), 8.13 (d, J=7.9 Hz, 1H), 8.02 (d,J=9.4 Hz, 1H), 7.88 (dd, J=4.7, 8.0 Hz, 1H), 7.84-7.49 (m, 3H), 7.15 (d,J=2.7 Hz, 1H), 5.82 (s, 1H), 3.38-3.19 (m, 2H), 3.14-3.06 (m, 1H),2.36-2.28 (m, 3H), 1.70-1.59 (m, 4H), 1.25-1.01 (m, 8H), additionalsignals missing due to water signal suppression; FXR EC₅₀=43 nM.

Example 1896-(2-(3-(3-Chloropyridin-2-yl)-5-cyclopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-N-(cyclopropylsulfonyl)-4-(difluoromethoxy)quinoline-2-carboxamide

The title compound was prepared as described in General Method E for thepreparation of Example 188 with replacement of4-(trifluoromethyl)nicotinaldehyde with 3-chloropicolinaldehyde. MS(ESI) m/z: 682.0 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ 8.68 (d, J=4.6 Hz,1H), 8.14 (d, J=8.2 Hz, 1H), 7.98 (d, J=9.2 Hz, 1H), 7.84-7.45 (m, 4H),7.15 (s, 1H), 5.88 (s, 1H), 3.26-3.16 (m, 2H), 3.04 (br s, 1H),2.38-2.28 (m, 3H), 1.73-1.61 (m, 4H), 1.26-0.87 (m, 8H), additionalsignals missing due to water signal suppression; FXR EC₅₀=48 nM.

6-(2-(3-(3-Chloropyridin-2-yl)-5-cyclopropylisoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(difluoromethoxy)-N-(methylsulfonyl)quinoline-2-carboxamide

The title compound was prepared as described in General Method E for thepreparation of Example 189 with replacement of cyclopropanesulfonamidewith methanesulfonamide. MS (ESI) m/z: 655.8 [M+H]⁺; ¹H NMR (500 MHz,DMSO-d₆) δ 8.69 (d, J=4.6 Hz, 1H), 8.15 (d, J=8.2 Hz, 1H), 8.02 (d,J=9.5 Hz, 1H), 7.90-7.55 (m, 4H), 7.16 (d, J=2.7 Hz, 1H), 5.89 (s, 1H),3.61-3.53 (m, 2H), 3.38 (s, 3H), 3.30-3.22 (m, 2H), 2.39-2.29 (m, 3H),1.74-1.61 (m, 4H), 1.23-1.10 (m, 4H); FXR EC₅₀=168 nM.

Example 1916-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)pyridin-3-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-N-(cyclopropylsulfonyl)-4-(difluoromethoxy)quinoline-2-carboxamide

The title compound was prepared as described in General Method E for thepreparation of Example 188 with replacement of4-(trifluoromethyl)nicotinaldehyde with2-(trifluoromethyl)nicotinaldehyde. MS (ESI) m/z: 716.3 [M+H]⁺; ¹H NMR(500 MHz, DMSO-d₆) δ 9.04 (d, J=5.2 Hz, 1H), 8.87 (s, 1H), 8.04-7.96 (m,2H), 7.88-7.54 (m, 3H), 7.15 (d, J=2.7 Hz, 1H), 5.83 (s, 1H), 3.27-3.18(m, 2H), 3.09 (dq, J=3.6, 4.2, 8.1 Hz, 1H), 2.39-2.29 (m, 3H), 1.73-1.58(m, 4H), 1.26-0.97 (m, 8H), additional signals missing due to watersignal suppression; FXR EC₅₀=140 nM.

Example 1926-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)pyridin-3-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(difluoromethoxy)-N-(methylsulfonyl)quinoline-2-carboxamide

The title compound was prepared as described in General Method E for thepreparation of Example 191 with replacement of cyclopropanesulfonamidewith methanesulfonamide. MS (ESI) m/z: 690.1 [M+H]⁺; ¹H NMR (500 MHz,DMSO-d₆) δ 9.03 (d, J=5.2 Hz, 1H), 8.84 (s, 1H), 8.01-7.90 (m, 2H),7.77-7.42 (m, 3H), 7.13 (d, J=2.7 Hz, 1H), 5.81 (d, J=2.4 Hz, 1H),3.51-3.40 (m, 2H), 3.19-3.09 (m, 2H), 2.96 (s, 3H), 2.39-2.27 (m, 3H),1.74-1.57 (m, 4H), 1.26-1.07 (m, 4H); FXR EC₅₀=238 nM.

Example 1936-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-N-(cyclopropylsulfonyl)quinoline-2-carboxamide

The title compound was prepared as described for the preparation ofExample 179 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl6-bromoquinoline-2-carboxylate. MS (ESI) m/z: 649.1 [M+H]⁺; ¹H NMR (500MHz, DMSO-d₆) δ 8.26 (d, J=8.58 Hz, 1H), 7.94-8.05 (m, 2H), 7.73 (br d,J=9.51 Hz, 1H), 7.65-7.69 (m, 2H), 7.60 (dd, J=7.24, 8.92 Hz, 1H),7.18-7.33 (m, 1H), 5.76-6.04 (m, 1H), 3.55 (br d, J=12.79 Hz, 2H), 2.55(s, 2H), 2.27-2.41 (m, 2H), 1.56-1.74 (m, 4H), 1.05-1.27 (m, 8H)additional signals missing due to water signal suppression; FXR EC₅₀=178nM.

Example 1946-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-N-(methylsulfonyl)quinoline-2-carboxamide

The title compound was prepared as described in General Method E for thepreparation of Example 193 with replacement of cyclopropanesulfonamidewith methanesulfonamide. MS (ESI) m/z: 623.1 [M+H]⁺; ¹H NMR (500 MHz,DMSO-d₆) δ 8.28 (d, J=8.54 Hz, 1H), 7.94-8.07 (m, 2H), 7.75 (dd, J=2.14,9.46 Hz, 1H), 7.65-7.71 (m, 2H), 7.57-7.64 (m, 1H), 7.26 (d, J=2.14 Hz,1H), 5.88 (s, 1H), 3.47-3.69 (m, 2H), 3.06-3.39 (m, 2H), 2.56 (s, 3H),2.37 (m, 3H), 1.53-1.84 (m, 4H), 0.93-1.29 (m, 4H); FXR EC₅₀=255 nM.

Example 1956-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-methoxy-N-(methylsulfonyl)quinoline-2-carboxamide

The title compound was prepared as described in General Method E for thepreparation of Example 184 with replacement of ethyl6-chloro-4-(trifluoromethyl)quinoline-2-carboxylate with methyl6-bromo-4-methoxyquinoline-2-carboxylate. MS (ESI) m/z: 653.9 [M+H]⁺; ¹HNMR (500 MHz, Methanol-d₄) δ 8.73 (s, 2H), 8.15 (d, J=9.35 Hz, 1H),7.83-7.98 (m, 1H), 7.77 (s, 1H), 7.65-7.74 (m, 1H), 5.85-6.07 (m, 1H),4.28 (s, 3H), 3.59-3.80 (m, 2H), 3.47 (br d, J=13.20 Hz, 2H), 3.39 (s,3H), 2.54 (s, 2H), 2.34 (s, 1H), 1.74-2.02 (m, 4H), 1.19-1.32 (m, 4H);FXR EC₅₀=75 nM.

Example 1966-(2-(5-Cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-methoxy-N-(methylsulfonyl)quinoline-2-carboxamide

The title compound was prepared as described in General Method E for thepreparation of Example 195 with replacement of3,5-dichloroisonicotinaldehyde with 2-(trifluoromethyl)benzaldehyde. MS(ESI) m/z: 653.1 [M+H]⁺; ¹H NMR (500 MHz, Methanol-d₄) δ 8.08 (d, J=9.35Hz, 1H), 7.89 (br d, J=7.15 Hz, 1H), 7.69-7.83 (m, 4H), 7.42-7.57 (m,2H), 5.63-5.98 (m, 1H), 4.24 (s, 3H), 3.51-3.63 (m, 2H), 3.33-3.38 (m,2H), 3.35 (s, 3H), 2.44 (s, 2H), 2.31 (m, 1H), 1.79 (m, 4H), 1.21 (m,4H); FXR EC₅₀=155 nM.

Example 1972-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluoro-N-sulfamoylbenzo[d]thiazole-6-carboxamide

The title compound was prepared as described in General Method E for thepreparation of Example 181 with replacement of cyclopropanesulfonamidewith sulfuric diamide. MS (ESI) m/z: 649.0 [M+H]⁺; ¹H NMR (500 MHz,CDCl₃) δ 8.58 (s, 2H), 7.92 (s, 1H), 7.53 (br d, J=11.28 Hz, 1H), 5.77(s, 1H), 3.66-3.84 (m, 2H), 3.44-3.59 (m, 2H), 2.40 (s, 2H), 2.07-2.21(m, 1H), 1.72 (br s, 4H), 1.21-1.34 (m, 2H), 1.16 (br d, J=6.05 Hz, 2H);FXR EC₅₀=381 nM.

General Method F Example 1982-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluoro-N-methylbenzo[d]thiazole-6-carboxamide

2-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid (15 mg, 0.03 mmol) was dissolved in DCE (1 mL).2,4,6-Tripropyl-1,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide (T3P)(0.03 mL, 0.05 mmol) followed by methylamine (1.6 mg, 0.05 mmol) andpyridine (6.4 μL, 0.08 mmol) were added to the reaction mixture and theresulting solution was stirred at room temperature for 6 h. The crudereaction mixture was directly purified flash chromatography on SiO₂(0-100% EtOAc/hexanes, Isco 12 g column) to yield2-(2-(5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluoro-N-methylbenzo[d]thiazole-6-carboxamide(10 mg, 0.016 mmol, 62% yield) as a white solid. MS (ESI) m/z: 584.0[M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δ 8.63 (s, 2H), 7.85 (d, J=1.54 Hz, 1H),7.38 (dd, J=1.54, 11.22 Hz, 1H), 6.04 (br d, J=4.62 Hz, 1H), 5.81 (s,1H), 3.65-3.88 (m, 2H), 3.42-3.60 (m, 2H), 3.01 (d, J=4.84 Hz, 3H), 2.44(s, 2H), 2.06-2.24 (m, 1H), 1.66-1.89 (m, 4H), 1.32 (dd, J=2.53, 4.95Hz, 2H), 1.07-1.21 (m, 2H); FXR EC₅₀=311 nM.

Example 1992-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluoro-N-isopropylbenzo[d]thiazole-6-carboxamide

The title compound was prepared as described in General Method F for thepreparation of Example 198 with replacement of methylamine withisopropylamine. MS (ESI) m/z: 612.0 [M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δ8.63 (s, 2H), 7.76-7.95 (m, 1H), 7.38 (d, J=11.22 Hz, 1H), 5.81 (s, 1H),4.29 (m, 2H), 3.64-3.93 (m, 2H), 3.53 (m, 1H), 2.44 (s, 2H), 2.18 (s,1H), 1.73-1.94 (m, 4H), 1.32 (m, 2H), 1.26 (d, J=6.60 Hz, 6H), 1.20 (m,2H); FXR EC₅₀=431 nM.

Example 2002-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-N-ethyl-4-fluorobenzo[d]thiazole-6-carboxamide

The title compound was prepared as described in General Method F for thepreparation of Example 198 with replacement of methylamine withethylamine. MS (ESI) m/z: 598.0 [M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δ 8.56(s, 2H), 7.78 (d, J=1.76 Hz, 1H), 7.33 (dd, J=1.54, 11.22 Hz, 1H), 6.02(s, 1H), 5.74 (s, 1H), 3.60-3.80 (m, 2H), 3.30-3.56 (m, 5H), 2.37 (s,2H), 1.98-2.21 (m, 2H), 1.45-1.77 (m, 7H), 0.74-1.33 (m, 13H); FXREC₅₀=68 nM.

Example 201N-Cyclopropyl-2-(2-(5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxamide

The title compound was prepared as described in General Method F for thepreparation of Example 198 with replacement of methylamine withcyclopropylamine. MS (ESI) m/z: 610.0 [M+H]⁺; ¹H NMR (400 MHz,CHLOROFORM-d) δ 8.63 (s, 2H), 7.83 (d, J=1.54 Hz, 1H), 7.36 (dd, J=1.54,11.22 Hz, 1H), 6.20 (br d, J=2.64 Hz, 1H), 3.76 (td, J=5.06, 13.20 Hz,2H), 3.45-3.64 (m, 2H), 2.89 (dt, J=3.30, 6.93 Hz, 1H), 2.44 (s, 2H),2.18 (tt, J=5.03, 8.39 Hz, 1H), 1.69-1.81 (m, 4H), 1.31 (dd, J=2.53,4.95 Hz, 2H), 1.13-1.24 (m, 2H), 0.87 (br d, J=5.50 Hz, 2H), 0.55-0.69(m, 2H); FXR EC₅₀=150 nM.

Example 2022-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxamide

The title compound was prepared as described in General Method F for thepreparation of Example 198 with replacement of methylamine with ammoniumchloride. MS (ESI) m/z: 570.0 [M+H]⁺; ¹H NMR (400 MHz, Methanol-d₄) δ8.63 (s, 2H), 7.92 (d, J=1.76 Hz, 1H), 7.50 (dd, J=1.54, 11.66 Hz, 1H),5.72-5.99 (m, 1H), 3.72 (br d, J=13.64 Hz, 2H), 3.43-3.57 (m, 2H), 2.41(s, 2H), 2.18-2.28 (m, 1H), 1.60-1.79 (m, 4H), 1.08-1.17 (m, 4H); FXREC₅₀=155 nM.

Example 203 Ethyl6-(2-(5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylate

A slurry of5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)-4-(7-azaspiro[3.5]non-1-en-2-yl)isoxazole(0.1 g, 0.27 mmol, synthesis described in General Method A), ethyl6-chloro-4-(trifluoromethyl)quinoline-2-carboxylate (0.097 g, 0.32 mmol)and Cs₂CO₃ (0.17 g, 0.53 mmol) in dioxane (1.8 mL) was degassed bybubbling nitrogen through the mixture for 5 min.Chloro(2-dicyclohexylphosphino-2′,6′-diisopropoxy-1,1′-biphenyl)[2-(2′-amino-1,1′-biphenyl)]palladium(II)(RuPhos-Pd-G2) (10.3 mg, 0.013 mmol) was then added and the reactionmixture was sealed and heated to 70° C. After heating for 3 h thereaction mixture was diluted with EtOAc, filtered and the filtrate wasconcentrated in vacuo. The residue was purified by flash chromatographyon SiO₂ (0-80% EtOAc/hexanes, Isco 40 g column) to give ethyl6-(2-(5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylate(61.7 mg, 0.095 mmol, 36% yield) as a red solid. MS (ESI) m/z: 643.1[M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δ 8.67 (s, 2H), 8.38 (s, 1H), 8.21 (d,J=9.7 Hz, 1H), 7.63 (dd, J=9.7, 2.6 Hz, 1H), 7.25 (br s, 1H), 5.86 (s,1H), 4.57 (q, J=7.0 Hz, 2H), 3.66-3.53 (m, 2H), 3.34 (ddd, J=12.8, 8.6,4.3 Hz, 2H), 2.47 (s, 2H), 2.29-2.18 (m, 2H), 1.87-1.77 (m, 4H), 1.50(t, J=7.2 Hz, 3H), 1.39-1.31 (m, 2H), 1.26-1.18 (m, 2H); FXR EC₅₀=946nM.

General Method G Example 2042-(2-(4-Cyclopropyl-1-(2,6-dichlorophenyl)-1H-1,2,3-triazol-5-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

Step 1. (2,6-dichlorophenyl)hydrazine

To a solution of 2,6-dichloroaniline (5.0 g, 30.9 mmol) in TFA (50 mL)was added water (10 mL). The reaction mixture was cooled to 0° C. andsodium nitrite (2.1 g, 30.9 mmol) was added over 0.5 hours, followed bygradual addition of sodium azide (5.1 g, 78.0 mmol) dissolved in aminimal volume of water. The mixture was stirred at 0° C. for 10minutes, and allowed to warm to room temperature. After 2 hours, thereaction mixture was filtered, the solid was washed with water, airdried and collected. The was filtrate diluted with EtOAc, washed withsaturated aqueous NaHCO₃ and brine, dried over MgSO₄, filtered andconcentrated in vacuo. The residue was purified by flash chromatographyon SiO₂ (0-20% EtOAc/hexanes). The solid isolated previously and theproduct from chromatography were combined to afford2-azido-1,3-dichlorobenzene (5.6 g, 29.9 mmol, 97% yield) as a tansolid. ¹H NMR (500 MHz, CDCl₃) δ 7.31 (d, J=8.3 Hz, 2H), 7.06 (t, J=8.1Hz, 1H).

Step 2. 4-cyclopropyl-1-(2,6-dichlorophenyl)-5-iodo-1H-1,2,3-triazole

To a room temperature solution of 2-azido-1,3-dichlorobenzene (4.5 g,23.7 mmol) in THF (120 mL) was added potassium iodide (15.8 g, 95 mmol)and copper (II) perchlorate hexahydrate (15.8 g, 42.7 mmol). Thereaction mixture was stirred at 50° C. for 5 minutes, followed byaddition of DBU (3.9 mL, 26.1 mmol) and cyclopropylacetylene (2.3 mL,27.3 mmol). The resulting brown mixture was stirred at room temperatureovernight. The reaction mixture was filtered through a pad of SiO₂ andconcentrated to dryness in vacuo. The residue was purified by flashchromatography on SiO₂ (0-20% EtOAc/hexanes) to afford4-cyclopropyl-1-(2,6-dichlorophenyl)-5-iodo-1H-1,2,3-triazole (1.8 g,4.8 mmol, 20% yield) as a white solid. ¹H NMR (500 MHz, CDCl₃) δ7.51-7.55 (m, 2H), 7.45-7.50 (m, 1H), 1.85-1.94 (m, 1H), 1.14-1.21 (m,2H), 1.03-1.10 (m, 2H).

Step 3. tert-Butyl2-(4-cyclopropyl-1-(2,6-dichlorophenyl)-1H-1,2,3-triazol-5-yl)-2-hydroxy-7-azaspiro[3.5]nonane-7-carboxylate

n-Butyllithium (2.5 M in hexanes, 0.26 mL, 0.66 mmol) was added slowlyto a −78° C. solution of4-cyclopropyl-1-(2,6-dichlorophenyl)-5-iodo-1H-1,2,3-triazole (0.2 g,0.53 mmol) in THF (2.1 mL) giving a dark brown solution. After 5minutes, a solution of tert-butyl2-oxo-7-azaspiro[3.5]nonane-7-carboxylate (0.15 g, 0.63 mmol) in 0.25 mLof THF was added slowly via syringe. The reaction was continued at −78°C. for 2 hours and brought to 0° C. for 45 minutes. The reaction wasquenched by the slow addition of approximately 1 mL of MeOH and thenconcentrated onto SiO₂ for purification. The residue was purified byflash chromatography on SiO₂ (0-100% EtOAc/hexanes, Isco 24 g column) togive tert-butyl2-(4-cyclopropyl-1-(2,6-dichlorophenyl)-1H-1,2,3-triazol-5-yl)-2-hydroxy-7-azaspiro[3.5]nonane-7-carboxylate(75 mg, 0.15 mmol, 29% yield) as a white foam.

Step 4.2-(4-Cyclopropyl-1-(2,6-dichlorophenyl)-1H-1,2,3-triazol-5-yl)-7-azaspiro[3.5]nonan-2-ol

tert-Butyl2-(4-cyclopropyl-1-(2,6-dichlorophenyl)-1H-1,2,3-triazol-5-yl)-2-hydroxy-7-azaspiro[3.5]nonane-7-carboxylate(75 mg, 0.152 mmol) was taken up in TFA (117 μL, 1.520 mmol). After 1 hthe excess TFA was removed in vacuo. The solid was dried in vacuoovernight and then used directly in the next step. MS (ESI) m/z: 393.1[M+H]⁺.

Step 5. Ethyl2-(2-(4-cyclopropyl-1-(2,6-dichlorophenyl)-1H-1,2,3-triazol-5-yl)-2-hydroxy-7-azaspiro[3.5]nonan-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylate

Cesium carbonate (61.9 mg, 0.19 mmol) followed by ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate (34.7 mg, 0.11 mmol) wereadded to a room temperature solution of2-(4-cyclopropyl-1-(2,6-dichlorophenyl)-1H-1,2,3-triazol-5-yl)-7-azaspiro[3.5]nonan-2-ol,TFA (38.6 mg, 0.08 mmol) in DMA (0.22 mL) and the reaction mixture washeated to 90° C. After 2 hours of heating the crude reaction mixture wasloaded directly onto a 12 g Isco SiO₂ cartridge for purification byflash chromatography (0-100% EtOAc/hexanes, Isco 12 g column) to giveethyl2-(2-(4-cyclopropyl-1-(2,6-dichlorophenyl)-1H-1,2,3-triazol-5-yl)-2-hydroxy-7-azaspiro[3.5]nonan-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylate(48 mg, 0.08 mmol, 100% yield). MS (ESI) m/z: 616.2 [M+H]⁺.

Example 204.2-(2-(4-Cyclopropyl-1-(2,6-dichlorophenyl)-1H-1,2,3-triazol-5-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid Phosphorus(V) oxychloride (45.4 μL, 0.49 mmol) followed by Et₃N(22.6 μL, 0.16 mmol) were added to a vial containing ethyl2-(2-(4-cyclopropyl-1-(2,6-dichlorophenyl)-1H-1,2,3-triazol-5-yl)-2-hydroxy-7-azaspiro[3.5]nonan-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylate(intermediate 33, 10 mg, 0.02 mmol). The reaction mixture was heated to60° C. for 5 hours and concentrated in vacuo to remove excess POCl₃. Theresidue was dissolved in THF (133 μL), water (53.3 μL), MeOH (13.33 μL)and lithium hydroxide monohydrate (8.4 mg, 0.20 mmol) was added to themixture. The reaction vessel was sealed and heated to 80° C. and afterheating over the weekend was quenched with 1N HCl, diluted with MeOH,and filtered. The solution was purified via preparative LC/MS with thefollowing conditions: Column: XBridge C18, 19×200 mm, 5-μm particles;Mobile Phase A: 5:95 acetonitrile: water with 10-mM ammonium acetate;Mobile Phase B: 95:5 acetonitrile: water with 10-mM ammonium acetate;Gradient: 10-60% B over 19 minutes, then a 5-minute hold at 100% B;Flow: 20 mL/min. Fractions containing the desired product were combinedand dried via centrifugal evaporation to give2-(2-(4-cyclopropyl-1-(2,6-dichlorophenyl)-1H-1,2,3-triazol-5-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid (4.6 mg, 8.1 mmol, 40% yield). MS (ESI) m/z: 570.2 [M+H]⁺; ¹H NMR(500 MHz, DMSO-d₆) δ 8.19 (s, 1H), 7.85-7.77 (m, 2H), 7.75-7.67 (m, 1H),7.58 (br d, J=11.6 Hz, 1H), 6.22 (s, 1H), 3.73 (br d, J=13.4 Hz, 1H),3.53 (br d, J=11.9 Hz, 1H), 2.36 (s, 2H), 2.13-2.00 (m, 1H), 1.77-1.59(m, 4H), 1.10-1.02 (m, 2H), 0.99 (br d, J=2.7 Hz, 2H); FXR EC₅₀=26 nM.

General Method H Example 2052-((2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-2-hydroxyspiro[3,5]nonan-7-yl)oxy)-4-fluorobenzo[d]thiazole-6-carboxylicacid

Step 1. 7-(tert-Butyldimethylsilyloxy)spiro[3,5]nonan-2-one

A solution of tert-butyldimethylchlorosilane (0.28 g, 1.9 mmol) in DCM(3 mL) was slowly added to a 0° C. solution of7-hydroxyspiro[3.5]nonan-2-one (0.25 g, 1.6 mmol) and imidazole (0.22 g,3.2 mmol) in DCM (5 mL). The ice bath was removed and the reactionmixture was stirred at room temperature overnight. The reaction mixturewas concentrated in vacuo and the residue was partitioned between EtOAcand saturated aqueous NaHCO₃. The organic phase was isolated, washedwith brine, dried over MgSO₄, filtered and concentrated in vacuo to give7-((tert-butyldimethylsilyl)oxy)spiro[3.5]nonan-2-one (0.408 g, 1.520mmol, 94% yield) as a colorless oil. MS (ESI) m/z: 269.2 [M+H]⁺; ¹H NMR(500 MHz, CDCl₃) δ 3.90-3.56 (m, 1H), 2.75 (br d, J=5.5 Hz, 4H),1.94-1.80 (m, 2H), 1.77-1.64 (m, 2H), 1.62-1.40 (m, 4H), 0.90 (s, 9H),0.07 (s, 6H).

Step 2.7-((tert-Butyldimethylsilyl)oxy)-2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)spiro[3.5]nonan-2-ol

n-Butyllithium (0.74 mL, 1.8 mmol) was added slowly to a −78° C.solution of 4-bromo-5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazole (0.49g, 1.5 mmol, synthesis described in General Method A) in THF (5.9 mL)giving a light brown solution. After 10 minutes,7-((tert-butyldimethylsilyl)oxy)spiro[3.5]nonan-2-one (0.40 g, 1.5 mmol)was added as a solution in ˜3 mL of THF. The reaction was continued at−78° C., and after 30 minutes, was quenched by the slow addition of 5 mLof MeOH and then concentrated in vacuo. The resulting residue waspurified by flash chromatography on SiO₂ (0-30% EtOAc/hexanes, 40 g IscoSiO₂ cartridge) to give7-((tert-butyldimethylsilyl)oxy)-2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)spiro[3.5]nonan-2-ol(0.48 g, 0.92 mmol, 62% yield) as a white foam. ¹H NMR (500 MHz, CDCl₃)δ 7.47-7.41 (m, 2H), 7.40-7.34 (m, 1H), 3.61-3.47 (m, 1H), 2.25 (s, 1H),2.21-2.08 (m, 3H), 2.04-1.88 (m, 3H), 1.60 (br d, J=12.1 Hz, 2H),1.45-1.36 (m, 1H), 1.34-1.18 (m, 6H), 1.17-1.08 (m, 2H), 0.87 (s, 9H),0.03 (s, 6H).

Step 3.2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)spiro[3.5]nonane-2,7-diol

To a room temperature solution of7-((tert-butyldimethylsilyl)oxy)-2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)spiro[3.5]nonan-2-ol(0.23 g, 0.43 mmol) in THF (2 mL) was added tetrabutylammonium fluoride(1 M in THF, 0.86 mL, 0.86 mmol). The reaction mixture was stirredovernight, quenched with 1.5 M aqueous potassium phosphate, andextracted twice with EtOAc. The combined organic extracts were washedwith brine, dried over Na₂SO₄, filtered, and concentrated in vacuo. Theresidue was purified by flash chromatography on SiO₂ (24 g, 0-100%EtOAc/hexanes over 7 minutes, then hold at 100% for 5 minutes, Isco 24 gSiO₂ column) to give2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)spiro[3.5]nonane-2,7-diol(0.16 g, 0.39 mmol, 91% yield) as a white foam. ¹H NMR (500 MHz, CDCl₃)δ 7.47-7.42 (m, 2H), 7.41-7.35 (m, 1H), 3.57 (br s, 1H), 2.27 (s, 1H),2.22-2.09 (m, 3H), 2.04-1.92 (m, 3H), 1.80-1.63 (m, 2H), 1.52-1.50 (m,1H), 1.48-1.36 (m, 1H), 1.36-1.23 (m, 6H), 1.18-1.08 (m, 3H).

Step 4. Methyl2-(2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-2-hydroxyspiro[3,5]nonan-7-yloxy)-4-fluorobenzo[d]thiazole-6-carboxlate

A solution of 2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)spiro[3.5]nonane-2,7-diol (33 mg, 0.081 mmol) in anhydrous THF (1 mL) atroom temperature was added KOtBu (19.0 mg, 0.17 mmol). After 5 minutes,methyl 2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate (28.1 mg, 0.097mmol) was added and the reaction mixture was stirred at room temperaturefor 5 minutes. The reaction was quenched with saturated aqueous NH₄Cl,and the resulting mixture was extracted three times with EtOAc. Thecombined organic extracts were washed with brine, dried over Na₂SO₄,filtered and concentrated in vacuo. The residue was purified by flashchromatography on SiO₂ (0-50% EtOAc/hexanes, Isco 12 g column) to givemethyl2-((2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-2-hydroxyspiro[3.5]nonan-7-yl)oxy)-4-fluorobenzo[d]thiazole-6-carboxylate(33 mg, 0.053 mmol, 66% yield) as a white foam. MS (ESI) m/z: 617.2[M+H]⁺.

Example 205.2-((2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-2-hydroxyspiro[3,5]nonan-7-yl)oxy)-4-fluorobenzo[d]thiazole-6-carboxylicacid

To a mixture of methyl2-((2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-2-hydroxyspiro[3.5]nonan-7-yl)oxy)-4-fluorobenzo[d]thiazole-6-carboxylate(33 mg, 0.053 mmol) was added MeOH (0.1 mL), water (0.40 mL) and THF(0.50 mL), followed by lithium hydroxide monohydrate (9.1 mg, 0.22mmol). The reaction mixture was stirred at 70° C. for 30 min andconcentrated in vacuo to remove THF and MeOH. The reaction mixture wasneutralized with 1 N aq. HCl to ˜pH 4 and the resulting suspension wasextracted with three times with EtOAc. The combined EtOAc extracts wereconcentrated in vacuo and the crude material was purified viapreparative LC/MS with the following conditions: Column: XBridge C18,19×200 mm, 5-μm particles; Mobile Phase A: 5:95 acetonitrile: water with10-mM ammonium acetate; Mobile Phase B: 95:5 acetonitrile: water with10-mM ammonium acetate; Gradient: 20-100% B) over 19 minutes, then a5-minute hold at 100% B). The desired fractions were combined andconcentrated to give2-((2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-2-hydroxyspiro[3,5]nonan-7-yl)oxy)-4-fluorobenzo[d]thiazole-6-carboxylicacid (5 mg, 8.2 μmol, 15% yield) as an off-white solid. MS (ESI) m/z:603.2 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ 8.31 (br s, 1H), 7.72-7.65 (m,1H), 7.64-7.58 (m, 2H), 7.58-7.51 (m, 1H), 5.18-5.02 (m, 1H), 3.60-3.42(m, 1H), 2.38-2.30 (m, 1H), 2.17-2.06 (m, 2H), 2.00-1.85 (m, 3H),1.85-1.75 (m, 1H), 1.65-1.51 (m, 2H), 1.50-1.40 (m, 1H), 1.40-1.32 (m,1H), 1.31-1.17 (m, 2H), 1.17-1.02 (m, 4H); GAL-FXR EC₅₀=2618 nM.

Example 2063-(((2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-2-hydroxyspiro[3.5]nonan-7-yl)oxy)methyl)benzoicacid

The title compound was prepared as described in General Method H for thepreparation of Example 205 with replacement of methyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl3-(bromomethyl)benzoate. MS (ESI) m/z: 542.2 [M+H]⁺; ¹H NMR (500 MHz,DMSO-d₆) δ 7.90-7.78 (m, 2H), 7.65-7.59 (m, 2H), 7.58-7.52 (m, 1H), 7.50(br d, J=7.3 Hz, 1H), 7.46-7.40 (m, 1H), 5.26 (s, 1H), 4.49 (s, 2H),3.20-3.10 (m, 1H), 2.39-2.29 (m, 1H), 2.12-2.01 (m, 2H), 1.95-1.82 (m,3H), 1.73-1.55 (m, 2H), 1.41-1.17 (m, 4H), 1.16-1.03 (m, 5H); GAL-FXREC₅₀=4711 nM.

General Method I Example 2073-((2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-2-hydroxyspiro[3.5]nonan-7-yl)oxy)benzoicacid

Step 1. Methyl3-((2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-2-hydroxyspiro[3.5]nonan-7-yl)oxy)benzoate

2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)spiro[3.5]nonane-2,7-diol(30 mg, 0.073 mmol, synthesis described in General Method H), methyl3-hydroxybenzoate (12.3 mg, 0.081 mmol), Bu₃P (0.029 mL, 0.12 mmol) and1,1′-(azodicarbonyl)dipiperidine (29.7 mg, 0.12 mmol) were dissolved indry dioxane (0.3 mL) in a sealed vial. The reaction mixture was heatedwith stirring at 110° C. for two hours. After cooling to roomtemperature, the mixture was diluted with water and extracted threetimes with EtOAc. The combined organic extracts were washed with brine,dried over Na₂SO₄, filtered and concentrated in vacuo. The residue waspurified by flash chromatography on SiO₂ (0-50% EtOAc/hexanes, Isco 12 gcolumn) to give methyl3-((2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-2-hydroxyspiro[3.5]nonan-7-yl)oxy)benzoate(10 mg, 0.018 mmol, 25% yield) as a white foam. MS (ESI) m/z: 542.2[M+H]⁺.

Example 207.3-((2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-2-hydroxyspiro[3.5]nonan-7-yl)oxy)benzoicacid

The hydrolysis of methyl3-((2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-2-hydroxyspiro[3.5]nonan-7-yl)oxy)benzoatewas accomplished as described in General Method H for the preparation ofExample 206. MS (ESI) m/z: 528.1 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ7.64-7.58 (m, 2H), 7.57-7.51 (m, 1H), 7.46 (br d, J=7.9 Hz, 1H),7.39-7.32 (m, 2H), 7.12 (br d, J=6.4 Hz, 1H), 5.35 (s, 1H), 4.35-4.21(m, 1H), 2.42-2.27 (m, 1H), 2.16-2.02 (m, 2H), 1.96-1.83 (m, 3H),1.79-1.68 (m, 1H), 1.68-1.59 (m, 1H), 1.54-1.43 (m, 1H), 1.41-1.30 (m,2H), 1.30-1.20 (m, 2H), 1.15-1.02 (m, 4H); FXR EC₅₀=4473 nM.

4-((2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-2-hydroxyspiro[3.5]nonan-7-yl)oxy)benzoicacid

The title compound was prepared as described in General Method I for thepreparation of Example 207 with replacement of methyl 3-hydroxybenzoatewith ethyl 4-hydroxybenzoate. MS (ESI) m/z: 528.1 [M+H]⁺; ¹H NMR (500MHz, DMSO-d₆) δ 7.82 (br d, J=8.5 Hz, 2H), 7.63-7.60 (m, 2H), 7.58-7.52(m, 1H), 6.92 (br d, J=8.5 Hz, 2H), 5.33 (s, 1H), 4.32-4.21 (m, 1H),2.39-2.30 (m, 1H), 2.16-2.04 (m, 2H), 1.97-1.84 (m, 3H), 1.80-1.60 (m,2H), 1.55-1.44 (m, 1H), 1.43-1.31 (m, 2H), 1.30-1.18 (m, 2H), 1.15-1.03(m, 4H); GAL-FXR EC₅₀=6660 nM.

Example 2092-((2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)spiro[3.5]non-1-en-7-yl)oxy)-4-fluorobenzo[d]thiazole-6-carboxylicacid

Step 1.2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-8,11-dioxadispiro[3.2.4⁷.2⁴]tridecan-2-ol

n-Butyllithium (2.5 M in hexanes, 1.2 mL, 3.0 mmol) was added slowly toa −78° C. solution of4-bromo-5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazole (0.80 g, 2.4 mmol,synthesis described in General Method A) in THF (9.6 mL). After 10minutes, 8,11-dioxadispiro[3.2.4⁷.2⁴]tridecan-2-one (0.47 mg, 2.4 mmol)was added as a solution in ˜0.5 mL of THF. After 30 minutes the reactionwas quenched by the slow addition of approximately 5 mL of MeOH and thenconcentrated in vacuo to dryness. The resulting residue was purified byflash chromatography on SiO₂ (0-60% EtOAc/hexanes, Isco 40 g column) togive2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-8,11-dioxadispiro[3.2.4⁷.2⁴]tridecan-2-ol(0.76 g, 1.69 mmol, 70% yield) as a white foam.

Step 2.2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-2-hydroxyspiro[3.5]nonan-7-one

To a mixture of2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-8,11-dioxadispiro[3.2.4⁷.2⁴]tridecan-2-ol(0.40 g, 0.89 mmol) in MeOH (4 mL) and water (4 mL) at room temperaturewas added p-toluenesulfonic acid monohydrate (84 mg, 0.44 mmol). Thereaction mixture was stirred at room temperature for 16 h and additionalp-toluenesulfonic acid monohydrate (84 mg, 0.44 mmol) was added. Afterone hour, the reaction was quenched with 1M K₂HPO₄ (20 mL), andextracted twice with EtOAc. The combined organic extracts were washedwith brine, dried over Na₂SO₄, filtered and concentrated in vacuo. Theresidue was purified by flash chromatography on SiO₂ (0-100% EtOAc/hex,Isco 24 g column) to give 2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-2-hydroxyspiro[3.5]nonan-7-one (0.32 g, 0.79 mmol, 89%yield) as an off-white foam. ¹H NMR (400 MHz, CDCl₃) δ 7.48-7.43 (m,2H), 7.41-7.35 (m, 1H), 2.36-2.33 (m, 1H), 2.33-2.30 (m, 1H), 2.30-2.22(m, 2H), 2.21-2.12 (m, 5H), 2.10-2.02 (m, 2H), 1.75 (t, J=6.6 Hz, 2H),1.33-1.24 (m, 3H), 1.19-1.10 (m, 2H).

Step 3.2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)spiro[3.5]non-1-en-7-one

To a reaction flask containing2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-2-hydroxyspiro[3.5]nonan-7-one(0.37 g, 0.91 mmol) was added TFA (1 mL, 13.0 mmol). The reactionmixture was stirred at room temperature for 30 minutes and concentratedin vacuo. The residue was diluted with EtOAc, washed with 1M K₂HPO₄,brine, dried over Na₂SO₄, filtered, and concentrated in vacuo. Theresidue was purified by flash chromatography on SiO₂ (0-30%EtOAc/hexanes, Isco 24 g cartridge) to give2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)spiro[3.5]non-1-en-7-one(0.28 g, 0.73 mmol, 80% yield) as a white foam. ¹H NMR (400 MHz, CDCl₃)δ 7.45-7.40 (m, 2H), 7.39-7.33 (m, 1H), 5.82 (s, 1H), 2.47 (s, 2H),2.39-2.28 (m, 4H), 2.18 (tt, J=8.4, 5.0 Hz, 1H), 1.99-1.82 (m, 4H),1.34-1.28 (m, 2H), 1.21-1.12 (m, 2H).

Step 4.2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)spiro[3.5]non-1-en-7-ol

To a solution of 2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)spiro[3.5]non-1-en-7-one (165 mg, 0.42 mmol) in MeOH (2.1 mL) at 0° C.was added NaBH₄ (17.7 mg, 0.47 mmol) in several portions. The reactionmixture was stirred at 0° C. for 30 min and concentrated in vacuo. Theresidue was diluted with EtOAc, washed with H₂O and brine, dried overNa₂SO₄, filtered and concentrated in vacuo. The residue was purified byflash chromatography on SiO₂ (0-100% EtOAc/hexanes, Isco 12 g column) togive2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)spiro[3.5]non-1-en-7-ol(0.14 g, 0.36 mmol, 84% yield) as a white solid.

Example 209.2-((2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)spiro[3.5]non-1-en-7-yl)oxy)-4-fluorobenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method H for thepreparation of Example 205 with replacement of2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)spiro[3.5]nonane-2,7-diolwith2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)spiro[3.5]non-1-en-7-ol.MS (ESI) m/z: 585.1 [M+H]⁺; NMR represents 1:1 mixture of isomers. ¹HNMR (500 MHz, DMSO-d₆) δ 8.35 (s, 2H), 7.78-7.56 (m, 8H), 6.00 (s, 1H),5.77 (s, 1H), 5.29-5.11 (m, 2H), 2.99 (s, 2H), 2.38-2.19 (m, 4H),2.09-1.74 (m, 8H), 1.69-1.47 (m, 8H), 1.23-1.17 (m, 4H), 1.13-1.08 (m,4H); FXR EC₅₀=1281 nM.

Example 2106-((2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)spiro[3.5]non-1-en-7-yl)oxy)quinoline-2-carboxylicacid

Step 1. Methyl6-((2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)spiro[3.5]non-1-en-7-yl)oxy)quinoline-2-carboxylate

2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)spiro[3.5]non-1-en-7-ol)(32 mg 0.082 mmol), methyl 6-hydroxyquinoline-2-carboxylate (20 mg,0.098 mmol), 1,1′-(azodicarbonyl)dipiperidine (33 mg, 0.13 mmol) andBu₃P (32 μL, 0.13 mmol) were dissolved in dry dioxane (0.41 mL). Thereaction mixture was stirred overnight at 100° C. in a sealed pressurevial. After cooling to room temperature, water was added, and theresulting mixture was extracted twice with EtOAc. The combined organiclayers were washed with brine, dried over Na₂SO₄, filtered andconcentrated in vacuo. The residue was purified by flash chromatographyon SiO₂ (0-60% EtOAc/hexanes, 10 minute gradient, then hold at 60% for 5minutes, Isco 12 g cartridge) to give methyl6-((2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)spiro[3.5]non-1-en-7-yl)oxy)quinoline-2-carboxylate(24 mg, 0.042 mmol, 51% yield, mixture of diastereomers) as a colorlessoil.

Example 210.6-((2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)spiro[3.5]non-1-en-7-yl)oxy)quinoline-2-carboxylicacid

The title compound was prepared as described in General Method H for thepreparation of Example 205 with replacement of methyl2-((2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-2-hydroxyspiro[3.5]nonan-7-yl)oxy)-4-fluorobenzo[d]thiazole-6-carboxylatewith methyl6-((2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)spiro[3.5]non-1-en-7-yl)oxy)quinoline-2-carboxylate.MS (ESI) m/z: 560.9 [M+H]⁺; ¹H NMR represents 1:1 mixture. ¹H NMR (500MHz, DMSO-d₆) δ 8.41-8.32 (m, 2H), 8.03 (d, J=8.5 Hz, 4H), 7.70-7.56 (m,6H), 7.50-7.38 (m, 4H), 6.00 (s, 1H), 5.76 (s, 1H), 4.64-4.43 (m, 2H),3.16 (s, 1H), 2.88 (s, 1H), 2.35-2.29 (m, 2H), 2.27 (s, 1H), 2.21 (s,1H), 2.02-1.80 (m, 4H), 1.71-1.41 (m, 12H), 1.24-1.18 (m, 4H), 1.12-1.07(m, 4H); FXR EC₅₀=485 nM.

Example 2112-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-1,4-dioxa-8-azaspiro[4.5]decan-8-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

Step 1. 5-cyclopropyl-3-(2,6-dichlorophenyl)-4-vinylisoxazole

To a solution of methyltriphenylphosphonium bromide (1.1 g, 3.2 mmol) inTHF (7.5 mL) at 0° C. was added KOtBu (1M in THF, 3.8 mL, 3.8 mmol)dropwise over 10 minutes. The reaction mixture was stirred for 30minutes followed by addition of5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazole-4-carbaldehyde (0.6 g, 2.1mmol, synthesis described in General Method C). The reaction mixture wasstirred at 0° C. for 30 minutes and directly purified by flashchromatography on SiO₂ (0-25% EtOAc/hexanes) to afford5-cyclopropyl-3-(2,6-dichlorophenyl)-4-vinylisoxazole (0.59 g, 2.1 mmol,99% yield) as a white crystalline solid. ¹H NMR (500 MHz, CDCl₃) δ7.45-7.40 (m, 2H), 7.40-7.32 (m, 1H), 6.39 (dd, J=17.9, 11.6 Hz, 1H),5.11 (d, J=11.6 Hz, 1H), 5.07-4.98 (m, 1H), 2.21-2.04 (m, 1H), 1.32-1.24(m, 2H), 1.15 (br dd, J=8.1, 2.3 Hz, 2H).

Step 2.1-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)ethane-1,2-diol

To a 0° C. solution of5-cyclopropyl-3-(2,6-dichlorophenyl)-4-vinylisoxazole (0.50 g, 1.8 mmol)in THF (5.3 mL) and water (5.3 mL) was added 4-methylmorpholine n-oxide(0.31 g, 2.7 mmol) and then osmium tetroxide (2.5% in tBuOH, 0.36 mL,0.036 mmol). The reaction mixture was stirred at 0° C. for 2 hours andthen allowed to warm to room temperature. After stirring overnight thereaction mixture was diluted with EtOAc, and washed with water. Theorganic was concentrated to a crude solid, which was then purified byflash chromatography on SiO₂ (0-75% EtOAc/hexanes) to afford1-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)ethane-1,2-diol(0.49 g, 1.5 mmol, 87% yield) as an off-white crystalline solid. ¹H NMR(500 MHz, CDCl₃) δ 7.40-7.45 (m, 2H), 7.33-7.39 (m, 1H), 4.49-4.68 (m,1H), 3.71-3.78 (m, 1H), 3.61-3.69 (m, 1H), 2.42 (br d, J=1.93 Hz, 1H),2.34 (ddd, J=3.58, 5.02, 8.46 Hz, 1H), 2.05 (s, 1H), 1.29-1.36 (m, 1H),1.21-1.28 (m, 1H), 1.09-1.17 (m, 2H).

Step 3. Ethyl4-fluoro-2-(4-oxopiperidin-1-yl)benzo[d]thiazole-6-carboxylate

A mixture of piperidin-4-one (48.9 mg, 0.49 mmol), ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate (150 mg, 0.49 mmol) andCs₂CO₃ (402 mg, 1.2 mmol) in DMF (1.5 mL) was heated at 60° C. for 1hour. The reaction mixture was diluted with EtOAc and washed with water.The organic layer was concentrated in vacuo and the residue was purifiedby flash chromatography on SiO₂ (0-60% EtOAc/hexanes) to afford ethyl4-fluoro-2-(4-oxopiperidin-1-yl)benzo[d]thiazole-6-carboxylate (41 mg,0.13 mmol, 26% yield) as a tan solid. ¹H NMR (500 MHz, CDCl₃) δ 8.14 (d,J=0.8 Hz, 1H), 7.80-7.68 (m, 1H), 4.46-4.32 (m, 2H), 4.09-3.94 (m, 4H),2.67 (t, J=6.3 Hz, 4H), 1.47-1.34 (m, 3H).

Step 4. Ethyl2-(2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-1,4-dioxa-8-azaspiro[4.5]decan-8-yl)-4-fluorobenzo[d]thiazole-6-carboxylate

To a mixture of1-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)ethane-1,2-diol(20.0 mg, 0.064 mmol, from step 2) and ethyl4-fluoro-2-(4-oxopiperidin-1-yl)benzo[d]thiazole-6-carboxylate (20.5 mg,0.064 mmol, from step 3) in DCE (0.5 mL) was added p-toluenesulfonicacid monohydrate (24.2 mg, 0.13 mmol). The reaction mixture was stirredat room temperature overnight. The reaction mixture was directlypurified by flash chromatography on SiO₂ (0-30% EtOAc/hex) to affordethyl2-(2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-1,4-dioxa-8-azaspiro[4.5]decan-8-yl)-4-fluorobenzo[d]thiazole-6-carboxylate(19.0 mg, 0.031 mmol, 48% yield) as a tan solid. ¹H NMR (500 MHz, CDCl₃)δ 8.11-8.07 (m, 1H), 7.72 (dd, J=11.3, 1.4 Hz, 1H), 7.47-7.41 (m, 2H),7.40 (d, J=8.0 Hz, 1H), 5.31 (s, 1H), 5.09-5.03 (m, 1H), 4.41-4.34 (m,2H), 4.16 (dd, J=8.3, 6.1 Hz, 1H), 3.86-3.67 (m, 4H), 3.60-3.52 (m, 1H),2.21-2.15 (m, 1H), 1.87-1.81 (m, 2H), 1.44-1.37 (m, 3H), 1.30 (br dd,J=5.0, 2.2 Hz, 3H), 1.20-1.13 (m, 2H).

Example 211.2-(2-(5-Cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-1,4-dioxa-8-azaspiro[4.5]decan-8-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

The hydrolysis of ethyl2-(2-(5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)-1,4-dioxa-8-azaspiro[4.5]decan-8-yl)-4-fluorobenzo[d]thiazole-6-carboxylateto give the title compound was accomplished as described in GeneralMethod C for the preparation of Example 39. MS (ESI) m/z: 590.2 [M+H]⁺;¹H NMR (500 MHz, DMSO-d₆) δ 8.12-8.21 (m, 1H), 7.64-7.68 (m, 2H),7.56-7.62 (m, 2H), 5.23 (br t, J=7.36 Hz, 1H), 4.24 (t, J=7.15 Hz, 1H),3.51 (br d, J=9.51 Hz, 1H), 3.21-3.40 (m, 1H), 2.55 (s, 2H), 2.34-2.40(m, 1H), 1.68-1.83 (m, 2H), 1.37 (br s, 1H), 1.17 (br d, J=8.25 Hz, 2H),1.09 (br s, 2H), 0.93-1.07 (m, 2H); FXR EC₅₀=1360 nM.

Example 2126-(2-(5-Cyclopropyl-3-(dicyclopropylmethyl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylicacid

The title compound was prepared as described for the preparation ofExample 82 with replacement of 2,6-difluorobenzaldehyde with2,2-dicyclopropylacetaldehyde. MS (ESI) m/z: 564.3 [M+H]⁺; ¹H NMR (500MHz, DMSO-d₆) δ 8.14 (s, 1H), 8.03 (br d, J=9.5 Hz, 1H), 7.83 (br d,J=9.5 Hz, 1H), 7.04 (br s, 1H), 6.37 (s, 1H), 3.60 (br s, 2H), 2.63 (s,2H), 2.49-2.46 (m, 2H), 2.14 (br s, 1H), 1.82 (br t, J=8.7 Hz, 1H), 1.70(br s, 4H), 1.15 (br s, 2H), 1.10 (br d, J=7.9 Hz, 2H), 1.01 (br d,J=7.9 Hz, 2H), 0.93 (br s, 2H), 0.43 (br d, J=3.7 Hz, 2H), 0.32-0.24 (m,2H), 0.21 (br dd, J=9.0, 4.4 Hz, 2H); FXR EC₅₀=1546 nM.

Example 2132-(2-(4-(Difluoromethyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

Step 1. Ethyl5-amino-1-(2-(trifluoromethyl)phenyl)-1H-pyrazole-4-carboxylate

A solution of ethyl (E)-2-cyano-3-ethoxyacrylate (1.6 g, 9.4 mmol) and(2-(trifluoromethyl)phenyl)hydrazine (1.5 g, 8.5 mmol) in ethanol (8.52mL) was heated to 85° C. in a sealed tube. Heating was continuedovernight, the reaction mixture was concentrated to minimal volume andthe residue was purified by flash chromatography on SiO₂ (0-100%EtOAc/hex, Isco 40 g column) to give ethyl5-amino-1-(2-(trifluoromethyl)phenyl)-1H-pyrazole-4-carboxylate (2.6 g,8.5 mmol, 100% yield as a brown solid. ¹H NMR (400 MHz, CDCl₃) δ 7.89(dd, J=7.6, 1.2 Hz, 1H), 7.82 (s, 1H), 7.78-7.71 (m, 1H), 7.71-7.64 (m,1H), 7.48 (d, J=7.7 Hz, 1H), 5.00 (br s, 2H), 4.32 (q, J=7.3 Hz, 2H),1.38 (t, J=7.2 Hz, 3H). ¹⁹F NMR (377 MHz, CDCl₃) δ −60.60 (s).

Step 2. Ethyl5-bromo-1-(2-(trifluoromethyl)phenyl)-1H-pyrazole-4-carboxylate

tert-Butyl nitrite (0.14 mL, 1.0 mmol) was added slowly to a roomtemperature suspension of copper(II) bromide (0.20 g, 0.92 mmol) andethyl 5-amino-1-(2-(trifluoromethyl)phenyl)-1H-pyrazole-4-carboxylate(0.25 g, 0.84 mmol) in acetonitrile (8.4 mL). After 1 h, the reactionwas quenched with saturated aqueous NaHCO₃. The mixture was taken up inEtOAc and washed with water and brine. The combined aqueous layers wereback extracted with EtOAc and the combined organics were dried overNa₂SO₄, filtered and concentrated to dryness in vacuo. The residue waspurified by flash chromatography on SiO₂ (0-50% EtOAc/hex, Isco 12 gcolumn) to give ethyl5-bromo-1-(2-(trifluoromethyl)phenyl)-1H-pyrazole-4-carboxylate (0.19 g,0.52 mmol, 62% yield) as a white solid. ¹H NMR (400 MHz, CDCl₃) δ 8.17(s, 1H), 7.92-7.86 (m, 1H), 7.80-7.69 (m, 2H), 7.43 (dd, J=7.4, 1.4 Hz,1H), 4.40 (q, J=7.3 Hz, 2H), 1.42 (t, J=7.2 Hz, 3H). ¹⁹F NMR (377 MHz,CDCl₃) δ −60.52 (s).

Step 3. tert-Butyl2-(4-(ethoxycarbonyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]non-1-ene-7-carboxylate

Tri-o-tolylphosphine (15.8 mg, 0.05 mmol), tert-butyl2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-7-azaspiro[3.5]non-1-ene-7-carboxylate(0.20 g, 0.57 mmol), ethyl5-bromo-1-(2-(trifluoromethyl)phenyl)-1H-pyrazole-4-carboxylate (0.19 g,0.52 mmol) and 2.0 M aqueous potassium phosphate (0.85 mL, 1.7 mmol)were dissolved in dioxane (3.3 mL) and the mixture was degassed bybubbling nitrogen through for 20 minutes. PdOAc₂ (5.8 mg, 0.03 mmol) wasadded and nitrogen was bubbled through the resulting mixture for 10minutes. The reaction vessel was sealed and heated to 80° C. After 3 hthe reaction mixture was diluted with EtOAc and washed with water andbrine. The combined aqueous layers were back extracted with EtOAc. Theorganic layers were dried over Na₂SO₄, filtered and concentrated todryness in vacuo. The residue was purified by flash chromatography onSiO₂ (0-80% EtOAc/hex, Isco 12 g column) to give tert-butyl2-(4-(ethoxycarbonyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]non-1-ene-7-carboxylate(0.24 g, 0.48 mmol, 92% yield) as a sticky solid. ¹H NMR (400 MHz,CDCl₃) δ 8.07 (s, 1H), 7.88-7.81 (m, 1H), 7.75-7.65 (m, 2H), 7.47-7.39(m, 1H), 6.39 (br s, 1H), 4.34 (q, J=7.0 Hz, 2H), 3.47 (dt, J=13.2, 5.1Hz, 2H), 3.20-3.09 (m, 2H), 2.18 (br s, 2H), 1.55-1.44 (m, 4H), 1.43 (s,9H), 1.40 (t, J=7.0 Hz, 3H). ¹⁹F NMR (377 MHz, CDCl₃) δ −60.51 (s, 1F).

Step 4. tert-Butyl2-(4-(hydroxymethyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]non-1-ene-7-carboxylate

Lithium aluminum hydride (2.5 mL, 2.5 mmol, 1M solution in THF) wasadded dropwise to a −50° C. solution of tert-butyl2-(4-(ethoxycarbonyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]non-1-ene-7-carboxylate (1.1 g,2.1 mmol) in THF (8.3 mL). The mixture was warmed to −10° C. for 35minutes and then brought to 0° C. for 40 minutes. The reaction wasquenched at 0° C. with sequential additions of 0.1 mL of water, 0.1 mLof 15% aqueous NaOH and 0.3 mL of water. The mixture was stirred at 0°C. for 10 minutes and at room temperature for 30 minutes. The layerswere separated and the organic layer was washed with water and brine.The aqueous layers were back extracted with EtOAc and the combinedorganics were dried over Na₂SO₄, filtered and concentrated to dryness invacuo. The residue was purified by flash chromatography on SiO₂ (0-100%EtOAc/hex, Isco 24 g column) to give tert-butyl2-(4-(hydroxymethyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]non-1-ene-7-carboxylate(0.78 g, 1.7 mmol, 81% yield) as an off-white foam. ¹H NMR (400 MHz,CDCl₃) δ 7.88-7.83 (m, 1H), 7.81 (s, 1H), 7.75-7.64 (m, 2H), 7.44-7.38(m, 1H), 4.64 (d, J=5.7 Hz, 2H), 1.59 (t, J=5.8 Hz, 1H). ¹⁹F NMR (377MHz, CDCl₃) δ −60.46 (s).

Step 5. tert-Butyl2-(4-formyl-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]non-1-ene-7-carboxylate

Triethylamine (97 μL, 0.69 mmol) followed byl-propanephosphonicanhydride (0.41 mL, 0.69 mmol, 50% solution in EtOAc) were added to a 0°C. solution of tert-butyl2-(4-(hydroxymethyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]non-1-ene-7-carboxylate(0.11 g, 0.23 mmol) in dichloromethane (1.3 mL and DMSO (1.0 mL). After20 minutes the reaction was quenched with brine and diluted with EtOAc.The aqueous layer was back extracted with EtOAc and the combinedorganics were dried over Na₂SO₄, filtered and concentrated to dryness invacuo. The residue was purified by flash chromatography on SiO₂ (0-100%EtOAc/hex, Isco 12 g column) to give tert-butyl2-(4-formyl-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]non-1-ene-7-carboxylate(0.085 g, 0.18 mmol, 80% yield) as a white foam. ¹H NMR (500 MHz, CDCl₃)δ 10.05 (s, 1H), 8.14 (s, 1H), 7.91-7.84 (m, 1H), 7.78-7.70 (m, 2H),7.49-7.44 (m, 1H), 6.27 (br d, J=2.2 Hz, 1H), 3.56-3.43 (m, 2H), 3.16(ddd, J=13.2, 8.9, 3.7 Hz, 2H), 2.22 (s, 2H), 1.44 (m, 13H).

Step 6. tert-Butyl2-(4-(difluoromethyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]non-1-ene-7-carboxylate

Diethylaminosulfur trifluoride (73.0 μL, 0.55 mmol) was added to a roomtemperature solution of tert-butyl2-(4-formyl-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]non-1-ene-7-carboxylate(85 mg, 0.18 mmol) in dichloromethane (1.8 mL). After stirring over theweekend, the reaction mixture was purified directly by flashchromatography on SiO₂ (0-80% EtOAc/hex, Isco 12 g column) to givetert-butyl2-(4-(difluoromethyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]non-1-ene-7-carboxylate(66 mg, 0.14 mmol, 74% yield) as a white foam. ¹H NMR (400 MHz, CDCl₃) δ7.91-7.83 (m, 2H), 7.78-7.68 (m, 2H), 7.51-7.45 (m, 1H), 6.98-6.61 (m,1H), 5.80 (s, 1H), 3.61-3.45 (m, 2H), 3.16 (ddd, J=13.2, 8.7, 4.1 Hz,2H), 2.21 (s, 2H), 1.55-1.48 (m, 4H), 1.46 (s, 9H).

Step 7.2-(4-(Difluoromethyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]non-1-ene

Trifluoroacetic acid (0.21 mL, 2.7 mmol) was added to a room temperaturesolution of tert-butyl2-(4-(difluoromethyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]non-1-ene-7-carboxylate(66 mg, 0.14 mmol) in dichloromethane (1.4 mL). After 2 hours thereaction mixture was concentrated to dryness to give2-(4-(difluoromethyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]non-1-ene,TFA. The product was used in subsequent steps without furtherpurification or characterization.

Example 213.2-(2-(4-(Difluoromethyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

A slurry of2-(4-(difluoromethyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]non-1-ene,TFA (30 mg, 0.06 mmol, from step 7), ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate (19 mg, 0.06 mmol) andCs₂CO₃ (59 mg, 0.18 mmol) in dioxane (0.3 mL) was heated to 80° C. in asealed flask. After 1.5 h, the reaction mixture was cooled to roomtemperature and THF/H₂O/MeOH (10:4:1, 0.6 mL) and LiOH monohydrate (13mg, 0.3 mmol) were added. The reaction mixture was sealed and heated to80° C. for 2 h. The reaction was quenched by the addition of 0.5 mL ofAcOH and concentrated to dryness in vacuo. The residue was dissolved in2 mL of MeOH, filtered, and the crude material was purified viapreparative LC/MS with the following conditions: Column: XBridge C18,200 mm×19 mm, 5-μm particles; Mobile Phase A: 5:95 acetonitrile: waterwith 10-mM ammonium acetate; Mobile Phase B: 95:5 acetonitrile: waterwith 10-mM ammonium acetate; Gradient: a 0-minute hold at 19% B, 19-59%B over 20 minutes, then a 4-minute hold at 100% B; Flow Rate: 20 mL/min;Column Temperature: 25° C. Fraction collection was triggered by MSsignals. Fractions containing the desired product were combined anddried via centrifugal evaporation to give2-(2-(4-(difluoromethyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid (23 mg, 0.04 mmol, 67% yield). MS (ESI) m/z: 579.3 [M+H]⁺; ¹H NMR(500 MHz, DMSO-d₆) δ 8.17 (s, 1H), 8.03-7.93 (m, 2H), 7.93-7.81 (m, 2H),7.66 (d, J=7.6 Hz, 1H), 7.57 (d, J=11.6 Hz, 1H), 7.30-7.02 (m, 1H), 5.95(s, 1H), 3.76-3.65 (m, 1H), 3.62-3.49 (m, 1H), 3.50-3.40 (m, 1H), 2.22(br s, 2H), 1.69-1.48 (m, 4H) additional peaks missing due to watersuppression; EC₅₀=48 nM.

Example 2146-(2-(4-(Difluoromethyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 82 with replacement of5-cyclopropyl-3-(2,6-difluorophenyl)-4-(7-azaspiro[3.5]non-1-en-2-yl)isoxazolewith2-(4-(difluoromethyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]non-1-ene.MS (ESI) m/z: 623.3 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ 8.20 (s, 1H),8.07 (d, J=9.4 Hz, 1H), 8.04-7.93 (m, 2H), 7.93-7.80 (m, 3H), 7.73-7.65(m, 1H), 7.33-7.02 (m, 2H), 5.98-5.92 (m, 1H), 3.33-3.22 (m, 1H), 2.23(br s, 2H), 1.72-1.62 (m, 2H), 1.60 (br s, 2H) additional peaks missingdue to water suppression; EC₅₀=26 nM.

Example 2152-(2-(4-(Difluoromethyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]nonan-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

Step 1.2-(4-(Difluoromethyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]nonane

A solution of tert-butyl2-(4-(difluoromethyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]non-1-ene-7-carboxylate(23.4 mg, 0.048 mmol) in MeOH (0.69 mL) was deoxygenated by bubblingnitrogen through the mixture for 5 minutes. To the mixture was addedpalladium on carbon (10% by wt., 25.8 mg, 0.024 mmol) and the reactionflask was sparged with hydrogen. A balloon of hydrogen was affixed andthe mixture was stirred at room temperature. After 2 h the reactionmixture was filtered through celite (EtOAc wash) and concentrated invacuo. The residue was dissolved in dichloromethane (0.47 mL) andtrifluoroacetic acid (46.8 μL) was added to the solution. After 1 h thesolvent was removed in vacuo and the resulting material was useddirectly in the next step.

Example 215.2-(2-(4-(Difluoromethyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]nonan-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 213 with replacement of2-(4-(difluoromethyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]non-1-enewith2-(4-(difluoromethyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]nonane.MS (ESI) m/z: 581.2 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ 8.16 (s, 1H),8.01-7.91 (m, 1H), 7.90-7.84 (m, 2H), 7.83-7.77 (m, 1H), 7.62 (br d,J=7.9 Hz, 1H), 7.56 (br d, J=11.3 Hz, 1H), 7.33-7.06 (m, 1H), 2.88 (s,1H), 2.72 (s, 1H), 2.11-2.02 (m, 1H), 1.97 (br t, J=10.4 Hz, 1H), 1.86(br d, J=8.2 Hz, 2H), 1.63 (br d, J=1.5 Hz, 2H), 1.50 (br s, 2H)Additional peaks lost under water peak; EC₅₀=995 nM.

Example 2162-(2-(1-(3,5-Dichloropyridin-4-yl)-4-(difluoromethyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 213 with replacement of(2-(trifluoromethyl)phenyl)hydrazine with3,5-dichloro-4-hydrazineylpyridine. MS (ESI) m/z: 580.1 [M+H]⁺; ¹H NMR(500 MHz, CDCl₃) δ 8.74 (s, 2H), 8.15 (s, 1H), 7.97 (s, 1H), 7.76 (br d,J=11.0 Hz, 1H), 7.01-6.62 (m, 1H), 6.03 (s, 1H), 3.91-3.78 (m, 2H), 3.74(s, 1H), 3.61-3.46 (m, 2H), 2.42 (s, 2H), 1.89-1.68 (m, 4H); EC₅₀=88 nM.

Example 2172-(2-(4-(Difluoromethyl)-1-(2-(trifluoromethoxy)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-fluorobenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 213 with replacement of(2-(trifluoromethyl)phenyl)hydrazine with(2-(trifluoromethoxy)phenyl)hydrazine. MS (ESI) m/z: 559.2 [M+H]⁺; ¹HNMR (500 MHz, DMSO-d₆) δ 8.16 (s, 1H), 7.98 (s, 1H), 7.74 (t, J=7.8 Hz,1H), 7.69 (d, J=7.6 Hz, 1H), 7.66-7.54 (m, 3H), 7.15 (t, J=55.0 Hz, 1H),6.06 (s, 1H), 3.67-3.44 (m, 4H), 2.31 (s, 2H), 1.63 (dt, J=4.9, 17.0 Hz,4H); EC₅₀=230 nM.

Example 2186-(2-(4-(Difluoromethyl)-1-(2-(trifluoromethoxy)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]non-1-en-7-yl)nicotinicacid

The title compound was prepared as described in General Method A for thepreparation of Example 217 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl6-fluoronicotinate. MS (ESI) m/z: 521.3 [M+H]⁺; ¹H NMR (500 MHz,DMSO-d₆) δ 8.58 (s, 1H), 7.99 (s, 1H), 7.89 (d, J=8.9 Hz, 1H), 7.78-7.68(m, 2H), 7.67-7.57 (m, 2H), 7.16 (t, J=55.1 Hz, 1H), 6.83 (d, J=9.2 Hz,1H), 6.04 (s, 1H), 3.86-7.78 (m, 2H), 3.45-3.36 (m, 2H), 2.29 (s, 2H),1.59-1.45 (m, 2H); EC₅₀=1085 nM.

Example 2196-(2-(4-(Difluoromethyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]non-1-en-7-yl)-1-methyl-1H-indole-3-carboxylicacid

The title compound was prepared as described in General Method B for thepreparation of Example 214 with replacement of ethyl6-bromo-4-(trifluoromethyl)quinoline-2-carboxylate with methyl6-bromo-1-methyl-1H-indole-3-carboxylate. MS (ESI) m/z: 557.3 [M+H]⁺; ¹HNMR (500 MHz, DMSO-d₆) δ 8.01 (d, J=7.5 Hz, 1H), 7.96 (s, 1H), 7.94-7.83(m, 2H), 7.81 (s, 1H), 7.77 (d, J=8.5 Hz, 1H), 7.68 (d, J=7.7 Hz, 1H),7.18 (t, J=55.1 Hz, 1H), 6.95-6.89 (m, 2H), 5.93 (s, 1H), 3.75 (s, 3H),3.27-3.19 (m, 2H), 2.91 (br t, J=10.4 Hz, 2H), 2.18 (s, 2H), 1.72-1.63(m, 2H), 1.60-1.53 (m, 2H); EC₅₀=180 nM.

Example 2202-(2-(4-(Difluoromethyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]nonan-7-yl)-4-methylbenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 215 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl2-bromo-4-methylbenzo[d]thiazole-6-carboxylate. MS (ESI) m/z: 577.2[M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ 8.12 (s, 1H), 7.98 (d, J=7.6 Hz,1H), 7.91-7.79 (m, 3H), 7.66 (s, 1H), 7.62 (d, J=7.8 Hz, 1H), 7.19 (t,J=55.6 Hz, 1H), 3.70-3.40 (m, 5H), 2.44 (s, 3H), 2.11-2.02 (m, 2H),2.01-1.92 (m, 2H), 1.86 (br d, J=9.3 Hz, 2H), 1.63 (br s, 2H), 1.50 (brs, 2H); EC₅₀=378 nM.

Example 2212-(2-(4-(Difluoromethyl)-1-(2-(trifluoromethoxy)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-methylbenzo[d]thiazole-6-carboxylicacid

The title compound was prepared as described in General Method A for thepreparation of Example 217 with replacement of ethyl2-bromo-4-fluorobenzo[d]thiazole-6-carboxylate with methyl2-bromo-4-methylbenzo[d]thiazole-6-carboxylate. MS (ESI) m/z: 591.1[M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ 8.14 (s, 1H), 7.99 (s, 1H),7.78-7.56 (m, 5H), 7.16 (t, J=55.0 Hz, 1H), 6.07 (s, 1H), 3.63-3.43 (m,4H), 2.45 (s, 3H), 2.32 (s, 2H), 1.71-1.56 (m, 4H); EC₅₀=182 nM.

Example 2226-(2-(4-(Difluoromethyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]nonan-7-yl)-1-methyl-1H-indole-3-carboxylic acid

The title compound was prepared as described in General Method B for thepreparation of Example 219 with replacement of2-(4-(difluoromethyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]non-1-ene,TFA with2-(4-(difluoromethyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]nonane,TFA. MS (ESI) m/z: 559.1 [M+H]⁺; ¹H NMR (500 MHz, DMSO-d₆) δ 7.98 (d,J=7.8 Hz, 1H), 7.91-7.85 (m, 2H), 7.84-7.79 (m, 2H), 7.76 (d, J=8.7 Hz,1H), 7.64 (d, J=7.8 Hz, 1H), 7.19 (t, J=55.5 Hz, 1H), 6.94-6.84 (m, 2H),3.73 (s, 2H), 3.03 (br s, 2H), 2.93 (br s, 2H), 2.07-1.99 (m, 2H),1.96-1.89 (m, 2H), 1.81 (br d, J=9.5 Hz, 2H), 1.65 (br s, 2H), 1.52 (brs, 2H), additional peak missing due to water suppression; EC₅₀=275 nM.

Example 2236-(2-(4-(Difluoromethyl)-1-(2-(trifluoromethoxy)phenyl)-1H-pyrazol-5-yl)-7-azaspiro[3.5]non-1-en-7-yl)nicotinicacid

The title compound was prepared as described in General Method A for thepreparation of Example 218 with replacement of(2-(trifluoromethoxy)phenyl)hydrazine with(2-(trifluoromethyl)phenyl)hydrazine. MS (ESI) m/z: 505.3 [M+H]⁺; ¹H NMR(500 MHz, DMSO-d₆) δ 8.56 (d, J=2.4 Hz, 1H), 7.98 (d, J=7.6 Hz, 1H),7.95 (s, 1H), 7.92-7.81 (m, 3H), 7.64 (d, J=7.6 Hz, 1H), 7.14 (t, J=55.0Hz, 1H), 6.80 (d, J=9.0 Hz, 1H), 5.91 (s, 1H), 3.83-3.74 (m, 2H),3.39-3.31 (m, 2H), 2.19 (s, 2H), 1.60-1.39 (m, 4H); EC₅₀=205 nM.

Example 224(2S,3S,4S,5R,6S)-6-((6-(2-(5-Cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carbonyl)oxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylicacid

Step 1.(2S,3R,4S,5S,6S)-6-((Allyloxy)carbonyl)-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl6-(2-(5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylate

To a mixture of6-(2-(5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylicacid (Example 85, 36.5 mg, 0.059 mmol) and HATU (24.8 mg, 0.065 mmol) inacetonitrile (0.5 mL) was added N-methylmorpholine (0.013 mL, 0.12 mmol)at room temperature. The reaction mixture was stirred at roomtemperature for 25 min followed by addition of allyl(2S,3S,4S,5R,6R)-3,4,5,6-tetrahydroxytetrahydro-2H-pyran-2-carboxylate(16.7 mg, 0.071 mmol). The resulting mixture was stirred at roomtemperature for 1.5 hours and purged with nitrogen to remove thesolvent. The residue was purified by flash chromatography on SiO₂(0-100% EtOAc/CH₂Cl₂, Isco 12 g column) to give(2S,3R,4S,5S,6S)-6-((allyloxy)carbonyl)-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl6-(2-(5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylate(31.8 mg, 0.038 mmol, 64% yield) as an orange solid. MS (ESI) m/z:831.5[M+H].

Step 2. Example 224

To a mixture of(2S,3R,4S,5S,6S)-6-((allyloxy)carbonyl)-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl6-(2-(5-cyclopropyl-3-(3,5-dichloropyridin-4-yl)isoxazol-4-yl)-7-azaspiro[3.5]non-1-en-7-yl)-4-(trifluoromethyl)quinoline-2-carboxylate(0.50 g, 0.60 mmol) and tetrakis(triphenylphosphine)palladium(0) (69.5mg, 0.060 mmol) in THF (8.0 mL) at 0° C. was added Et₃N (0.13 mL, 0.96mmol). After stirring at 0° C. for 40 min, acetonitrile was addedfollowed by Celite. The mixture was then evaporated to remove thesolvents, loaded into a cartridge and then purified by C-18 reversephase flash chromatography (100 g Isco HP C-18 column, 10-100% B in A,mobile phase A=10:90 acetonitrile: water with 0.05% trifluoroaceticacid; mobile phase B=90:10 acetonitrile: water with 0.05%trifluoroacetic acid). The collected fractions containing desiredproduct were purged with nitrogen to remove the solvents and the residuewas lyophilized to give(2S,3S,4S,5R,6S)-6-((6-(2-((5-cyclopropyl-3-(2-(trifluoromethyl)phenyl)isoxazol-4-yl)methyl)-7-azaspiro[3.5]nonan-7-yl)-4-(trifluoromethyl)quinoline-2-carbonyl)oxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylicacid (0.34 g, 0.43 mmol, 67% yield) as a light brown solid. MS (ESI)m/z: 791.2[M+H]⁺; 1H NMR (500 MHz, DMSO-d₆) δ 12.98-12.86 (m, 1H), 8.87(s, 2H), 8.26 (s, 1H), 8.11 (d, J=9.5 Hz, 1H), 7.89 (br dd, J=9.8, 2.5Hz, 1H), 7.06 (br s, 1H), 5.99 (s, 1H), 5.76 (d, J=7.6 Hz, 1H), 3.90 (d,J=9.5 Hz, 1H), 3.68-3.57 (m, 2H), 3.46-3.43 (m, 1H), 3.43-3.41 (m, 1H),3.40-3.38 (m, 1H), 3.33 (s, 2H), 2.41 (s, 2H), 2.37-2.33 (m, 1H),1.72-1.63 (m, 4H), 1.26-1.19 (m, 2H), 1.19-1.12 (m, 2H); FXR EC50=73 nM.

Biological Assays

The exemplified compounds of the present invention were tested in thetransient human FXR/Gal4-luciferase reporter assay, and assay resultswere reported in the Examples section with other analytical data.

A Gal4-hFXR fusion construct reporter system was used as the primaryassay to characterize compound activity. A construct including 5 copiesof the Gal4 promoter response element upstream of a firefly luciferasereporter cDNA was stably expressed in HEK293 cells. This reporter cellline was maintained in Dulbecco's Modified Eagle's medium (DMEM; Gibco)supplemented with 1% penicillin-streptomycin (P/S) solution, 500 μg/mLZeocin and 10% charcoal/dextran-treated fetal bovine serum (cs-FBS) at37° C. in a humidified 5% CO₂ atmosphere. Another plasmid wasconstructed in which the human cytomegalovirus promoter in the pcDNA3.1vector directs the expression of the cDNA encoding a fusion proteincomprised of the DNA binding domain from the Gal4 transcription factorfused to the ligand binding domain from human FXR.

The day prior to transfection, the reporter cells in culture aredetached from the plate with trypsin and plated into a T75 flask at asufficient density to achieve approximately 90% confluence the nextmorning. The transfection reagents are prepared by separately diluting25 μg of the pcDNA3.1-Gal4-FXR plasmid into 1.87 mL of Opti-MEM(Thermo-Fisher), and 40 μL of Lipofectamine 2000 (Thermo-Fisher) into1.87 mL of Opti-MEM, and then adding the diluted DNA solution into thediluted Lipofectamine 2000 solution and incubating at room temperaturefor 15-20 minutes. The mixture is further diluted with 10 mL of asolution comprised of DMEM, 10% cs-FBS, and 1% P/S immediately prior totransferring to the cells. The maintenance culture media is aspiratedfrom the cells and the final transfection mixture is added before thecells are incubated overnight at 37° C. in a humidified 5% CO₂atmosphere. This protocol can be scaled up, and the transientlytransfected cells can be cryopreserved in an assay-ready format.

For compound testing, 100 nL of the compounds (serial dilutions in DMSO)are dispensed with an Echo acoustic dispenser (Labcyte) into the wellsof a Corning/Costar clear bottom 384-well white plate. The transfectedcells are harvested, counted, and diluted such that 10-25,000 cells in25 μL are plated into each well of the 384-well compound assay plate.The compound-treated cells are incubated overnight at 37° C. in ahumidified 5% CO₂ atmosphere. The next morning 25 μL of Steady-Glo(Promega) are added to each well of the plate, the mixture is incubatedfor 15 min. with shaking, and luminescence is measured on an Envision(Perkin Elmer) plate reader. Background counts from cells treated withDMSO alone are subtracted from all raw counts, and the corrected valuesare converted to a percentage of the control response attained with 8 μMGW-4064. These data are fit to a 4-parameter log agonist-responseequation to calculate an EC₅₀ value.

Acute Mouse In Vivo Assay:

Male, C57BL6/NTac mice, weighing 25-28 g, are purchased from TaconicLabs (Hudson, N.Y.) and maintained on Teklad Global 18% Protein RodentDiet (Harlan Laboratories). After 1 week acclimation, mice are sortedinto groups based upon body weight. Mice are administered a single oraldose of vehicle or experimental compound. Systemic compound exposure isevaluated in plasma derived from blood collected via the submandibularvein at 1 hour post-dose, and at study termination (6 h). At studytermination, the animals are euthanized and rapidly dissected. Themedial lobe of the liver is divided, with one half being homogenized andanalyzed for compound exposure, and the other half saved in RNAlater(Thermo-Fisher Scientific). The ileum is also dissected and preserved inRNAlater. Tissue samples in RNAlater are homogenized with MPBiomedicals' beads. RNA is extracted using the MagMax-96 Total RNAIsolation kit (Thermo-Fisher Scientific) according to the manufacturer'sprotocol. RNA Concentration is determined with the Nano-Drop 8000Spectrophotometer (Thermo Fisher). Reverse transcription is done withInvitrogen's SuperScript® VILO cDNA Synthesis Kit according to themanufacturer's protocol. Real time PCR is done with Applied Biosystems'Taqman PCR master mixture according to the manufacturer's protocol. Allprimers are purchased from Thermo-Fisher Scientific. Mouse genesanalyzed include Nr0b2 (which encodes the small heterodimer partner,SHP), Abcb11 (which encodes the bile salt excretion pump, BSEP), Cyp7a1,& Cyp8b1 in liver, and Fgf15, Fabp6 (which encodes ileal bile acidbinding protein, I-BABP), Slc51a (which encodes organic solutetransporter alpha subunit, OSTA), and Slc51b (which encodes organicsolute transporter beta subunit, OSTB) in the ileum. The statisticalsignificant changes in FGF15 gene expression are expressed as foldincrease and CYP_(7A1) expression as a percent reduction relative tovehicle control.

Other features of the invention should become apparent in the course ofthe above descriptions of exemplary embodiments that are given forillustration of the invention and are not intended to be limitingthereof. The present invention may be embodied in other specific formswithout departing from the spirit or essential attributes thereof. Thisinvention encompasses all combinations of preferred aspects of theinvention noted herein. It is understood that any and all embodiments ofthe present invention may be taken in conjunction with any otherembodiment or embodiments to describe additional embodiments. It is alsounderstood that each individual element of the embodiments is its ownindependent embodiment. Furthermore, any element of an embodiment ismeant to be combined with any and all other elements from any embodimentto describe an additional embodiment.

What is claimed is:
 1. A compound of Formula (I):

or a stereoisomer, a tautomer, or a pharmaceutically acceptable salt orsolvate thereof; wherein: X¹ and X⁴ are each independently C or N; X²and X³ are each independently CR⁵, N, NR⁶, O, or S; E ring is a 4- to6-membered carbocyclyl or heterocyclyl, wherein the carbocyclyl andheterocyclyl are each independently substituted with 0 to 3 R³; *denotes a spiro carbon atom; Y is CR⁷ or N; m and n are eachindependently an integer of 0, 1, or 2; f is an integer of 0, 1, 2, or3; Z is 6- to 10-membered aryl, 5- to 10-membered heteroaryl containing1 to 3 heteroatoms independently selected from N, O, and S, 3- to10-membered carbocyclyl, or 4- to 10-membered heterocyclyl containing 1to 3 heteroatoms independently selected from N, O, and S, wherein thearyl, heteroaryl carbocyclyl, and heterocyclyl are independentlysubstituted with 0 to 5 R⁸; L¹ is a covalent bond, O, S, —NR¹⁶—,—S(O)₂—, C₁₋₃ alkylene, C₁₋₃ heteroalkylene, C₂₋₄ alkenylene, C₂₋₄alkynylene, aryl, or a 5- to 6-membered heteroaryl containing 1 to 3heteroatoms independently selected from N, O, and S containing 1 to 4heteroatoms independently selected from N, O, and S; wherein thealkylene, alkenylene, aryl, heteroalkylene, and heteroaryl are eachindependently substituted with 0 to 3 R¹; L² is a covalent bond, O, S,—NR¹⁷—, C₁₋₃ alkylene, or C₁₋₃ heteroalkylene, wherein the alkylene andheteroalkylene are independently substituted with 0 to 3 R¹⁵; R^(X) is-L³-R^(Z); L³ is a covalent bond, C₁₋₃ alkylene, —C(O)NR¹²—CH₂—, or—OCH₂—, wherein the C₁₋₃ alkylene is substituted with 0 to 3 R⁴; R^(Z)is —CN, —C(O)OR¹³, —C(O)NR^(14a)R^(14b),

R^(e) is C₁₋₆ alkyl, C₃₋₆ cycloalkyl, haloalkyl, hydroxyalkyl,aminoalkyl, alkoxyalkyl, haloalkoxyalkyl, or aryl; R^(Y) is eachindependently hydrogen, halo, cyano, hydroxyl, amino, C₁₋₆ alkyl,alkylamino, haloalkyl, hydroxyalkyl, aminoalkyl, alkoxyalkyl,haloalkoxyalkyl, alkoxy, or haloalkoxy; or alternatively two R^(Y),together with the carbon atoms to which they are attached, form a bridgemoiety; and with the proviso that when Y is N and R^(Y) is attached to acarbon atom adjacent to Y, then R^(Y) is not halo, cyano, hydroxyl,amino, alkoxy, or haloalkoxy; R¹ is C₁₋₆ alkyl, C₃₋₅ cycloalkyl, or C₄₋₆heterocyclyl, wherein the alkyl, cycloalkyl, and heterocyclyl are eachsubstituted with 0 to 3 R⁹; R² is 6- to 10-membered aryl, 5- to10-membered heteroaryl containing 1 to 3 heteroatoms independentlyselected from N, O, and S, 3- to 10-membered carbocyclyl, or 4- to10-membered heterocyclyl containing 1 to 3 heteroatoms independentlyselected from N, O, and S, wherein the aryl, heteroaryl, carbocyclyl,and heterocyclyl are independently substituted with 0 to 5 R¹⁰; R³, R⁵,and R⁷ are each independently hydrogen, halo, cyano, hydroxyl, amino,C₁₋₆ alkyl, alkylamino, haloalkyl, hydroxyalkyl, aminoalkyl,alkoxyalkyl, haloalkoxyalkyl, alkoxy, or haloalkoxy; R⁴ is eachindependently halo, oxo, cyano, hydroxyl, amino, alkyl, alkoxy, oralkylamino; or alternatively, two R⁴, taken together with the atom(s) towhich they are attached, form a carbocyclyl or heterocyclyl moiety; R⁶,R¹⁶ and R¹⁷ are each independently hydrogen, C₁₋₆ alkyl, haloalkyl,hydroxyalkyl, aminoalkyl, alkoxyalkyl, or haloalkoxyalkyl; R⁸ and R¹⁰are each independently halo, cyano, hydroxyl, amino, oxo, —OR^(a),—SR^(a), ═S, —NR^(c)R^(c), ═NH, ═N—OH, ═NR^(a), ═N—OR^(a), —NO₂,—S(O)₂R^(a), —S(O)₂NHR^(b), —S(O)₂NR^(c)R^(c), —S(O)₂OR^(b),—OS(O)₂R^(b), —OS(O)₂OR^(b), —P(O)(OR^(b))(OR^(b)), —C(O)R^(b),—C(NR^(b))R^(b), —C(O)OR^(b), —C(O)NR^(c)R^(c), —C(NR^(b))NR^(c)R^(c),—OC(O)R^(b), —NR^(b)C(O)R^(b), —OC(O)OR^(b), —NR^(b)C(O)OR^(b),—NR^(b)C(O)NR^(c)R^(c), —NR^(b)C(NR^(b))R^(b),—NR^(b)C(NR^(b))NR^(c)R^(c), C₁₋₆ alkyl, C₁₋₆ haloalkyl, aryl,arylalkyl, heteroaryl, carbocyclyl, or heterocyclyl; wherein the alkyl,aryl, heteroaryl, carbocyclyl, and heterocyclyl, by themselves or aspart of another group, are each independently substituted with 0 to 5R^(d); R^(a) is each independently C₁₋₆ alkyl, haloalkyl, hydroxyalkyl,aminoalkyl, alkoxyalkyl, haloalkoxyalkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl, carbocyclyl, carbocyclylalkyl, heterocyclyl, orheterocyclylalkyl; R^(b) is each independently hydrogen or R^(a); R^(c)is each independently R^(b) or alternatively, the two R^(c) are takentogether with the nitrogen atom to which they are bonded form a 4-, 5-,6- or 7-membered heterocyclyl containing 1 to 3 heteroatomsindependently selected from N, O, and S; R^(d) is each independentlyR^(a), alkoxy, haloalkoxy, alkylamino, cycloalkylamino,heterocyclylamino, haloalkyl, hydroxyalkyl, aminoalkyl, cycloalkoxy,heterocyclyloxy, haloalkoxy, alkoxyalkoxy, haloalkylamino,alkoxyalkylamino, haloalkoxyalkylamino, arylamino, aralkylamino,aryloxy, aralkyloxy, heteroaryloxy, heteroarylalkyloxy, alkylthio, halo,cyano, hydroxyl, amino, oxo, —OR^(a), —SR^(a), ═S, —NR^(c)R^(c), ═NH,═N—OH, ═NR^(a), ═N—OR^(a), —NO₂, —S(O)₂R^(a), —S(O)₂NHR^(b),—S(O)₂NR^(c)R^(c), —S(O)₂OR^(b), —OS(O)₂R^(b), —OS(O)₂OR^(b),—P(O)(OR^(b))(OR^(b)), —C(O)R^(b), —C(NR^(b))R^(b), —C(O)OR^(b),—C(O)NR^(c)R^(c), —C(NR^(b))NR^(c)R^(c), —OC(O)R^(b), —NR^(b)C(O)R^(b),—OC(O)OR^(b), —NR^(b)C(O)OR^(b), —NR^(b)C(O)NR^(c)R^(c),—NR^(b)C(NR^(b))R^(b), or —NR^(b)C(NR^(b))NR^(c)R^(c); R⁹ is eachindependently halo, cyano, hydroxyl, amino, or C₁₋₆ alkyl; R¹¹ and R¹⁵are each independently halo, oxo, cyano, hydroxyl, amino, C₁₋₆ alkyl,C₃₋₆ cycloalkyl, C₄₋₆ heterocyclyl, alkylamino, haloalkyl, hydroxyalkyl,aminoalkyl, alkoxyalkyl, haloalkoxyalkyl, alkoxy, or haloalkoxy; R¹² ishydrogen or C₁₋₄ alkyl; R¹³ is hydrogen, C₁₋₁₀ alkyl, glycosyl, orcarboxy(trihydroxy)tetrahydropyranyl; and R^(14a) and R^(14b) are eachindependently hydrogen, C₁₋₆ alkyl, C₃₋₆ cycloalkyl, C₄₋₆ heterocyclyl,alkylamino, haloalkyl, hydroxyalkyl, aminoalkyl, alkoxyalkyl,haloalkoxyalkyl, alkoxy, or haloalkoxy.
 2. The compound according toclaim 1, or a stereoisomer, a tautomer, or a pharmaceutically acceptablesalt or solvate thereof; wherein: the

moiety is


3. The compound according to claim 1, or a stereoisomer, a tautomer, ora pharmaceutically acceptable salt or solvate thereof; wherein: the

moiety is


4. The compound according to claim 1, or a stereoisomer, a tautomer, ora pharmaceutically acceptable salt or solvate thereof; wherein: L¹ is acovalent bond, O, S, NH, C₁₋₃ alkylene, —(C₁₋₃ alkylene)_(a)-O—(C₁₋₃alkylene)_(b)-, —(C₁₋₃ alkylene)_(a)-S—(C₁₋₃ alkylene)_(b)-, or —(C₁₋₃alkylene)_(a)-NH—(C₁₋₃ alkylene)_(b)-, wherein the C₁₋₃ alkylene issubstituted with 0 to 3 R¹; a is an integer of 0 or 1; b is an integerof 0 or 1; provided that a and b are not both 1; and L² is a covalentbond.
 5. The compound according to claim 1, or a stereoisomer, atautomer, or a pharmaceutically acceptable salt or solvate thereof;wherein: the

moiety is selected from:


6. The compound according to claim 1, or a stereoisomer, a tautomer, ora pharmaceutically acceptable salt or solvate thereof, wherein Z isphenyl or 5- to 10-membered heteroaryl containing 1 to 3 heteroatomsindependently selected from N, O, and S, wherein the phenyl andheteroaryl are independently substituted with 0 to 5 R⁸.
 7. The compoundaccording to claim 1, which is represented by Formula (III):

or a stereoisomer, a tautomer, or a pharmaceutically acceptable salt orsolvate thereof; wherein: Z is 6-membered monocyclic heteroarylcontaining 1 or 2 nitrogen atoms, or a 9- to 10-membered bicyclicheteroaryl containing 1 or 3 heteroatoms independently selected from N,O, and S, wherein the monocyclic or bicyclic heteroaryl is independentlysubstituted with 0 to 3 R⁸; R² is phenyl or pyridinyl, wherein thephenyl and pyridinyl are each independently substituted with 0 to 2 R¹⁰;R⁸ is each independently halo, cyano, hydroxyl, C₁₋₄ alkyl, C₁₋₄haloalkyl, C₁₋₄ alkoxy, or C₁₋₄ haloalkoxy; R¹⁰ is each independentlyhalo, C₁₋₄ alkyl, C₁₋₄ haloalkyl, C₁₋₄ alkoxy, or C₁₋₄ haloalkoxy; R^(X)is —C(O)OH or —C(O)NH—S(O)₂R^(e); and R^(e) is C₁₋₆ alkyl or C₃₋₆cycloalkyl.
 8. The compound according to claim 1, or a stereoisomer, atautomer, or a pharmaceutically acceptable salt or solvate thereof;wherein: X¹ is C, X² is N, X³ is O, and X⁴ is C; or X¹ is N, X² is N, X³is C, and X⁴ is C; Y is CH or N; the

moiety is selected from:

L¹ is a covalent bond, O, or —OCH₂—, provided that L¹ is a covalent bondwhen Y is N; Z is phenyl, pyridinyl, pyridazinyl, pyrimidinyl,pyrazinyl, benzo[d]imidazolyl, benzo[d]isoxazolyl, benzo[d]oxadiazolyl,benzo[d]thiazolyl, imidazolo[1,5-a]pyridinyl, indazolyl, indolyl,pyrazolo[4,3-b]pyridinyl, pyrrolo[2,1-f][1,2,4]triazinyl,pyrrolo[2,3-b]pyridinyl, pyrrolo[2,3-c]pyridinyl,pyrrolo[2,3-d]pyrimidinyl, pyrrolo[3,2-c]pyridinyl,thiazolo[4,5-b]pyridinyl, thiazolo[5,4-b]pyridinyl, cinnolinyl,isoquinolinyl, quinolinyl, or quinoxalinyl, each substituted with zeroto 1 R⁸; R⁸ is F, —CH₃, —CF₃, —OCH₃, —OCH₂CH₃, —OCH(CH₃)₂, —OCHF₂,—OCF₃, —OCH₂CH₂OH, or —CH₂OCH₂CH₂Si(CH₃)₃; R^(x) is —CN, —C(O)OH,—C(O)OCH₂CH₃, —C(O)NH₂, —C(O)NH(CH₃), —C(O)NHCH₂CH₃, —C(O)NHCH(CH₃)₂,—C(O)NH(cyclopropyl), —C(O)NHCH₂C(O)OH, —C(O)NHS(O)₂CH₃,—C(O)NHS(O)₂(cyclopropyl), —OCH₂C(O)OH, or—C(O)O(carboxy(trihydroxy)tetrahydropyranyl); L² is a covalent bond; R¹is —CHF₂, —CH(CH₃)₂, cyclopropyl, or methylcyclopropyl; R² iscyclohexyl, phenyl, or pyridinyl, wherein the phenyl and the pyridinylare independently substituted with 1 to 3 R¹⁰; and R¹⁰ is eachindependently F, C₁, —CH₃, —CF₃, —OCH₃, or —OCF₃.
 9. The compoundaccording to claim 1, or a stereoisomer, a tautomer, or apharmaceutically acceptable salt or solvate thereof, selected from:


10. The compound according to claim 1, or a stereoisomer, a tautomer, ora pharmaceutically acceptable salt or solvate thereof, selected from:


11. A pharmaceutical composition comprising a pharmaceuticallyacceptable carrier and a compound according to claim 1, or astereoisomer, a tautomer, or a pharmaceutically acceptable salt orsolvate thereof.
 12. A pharmaceutical composition comprising apharmaceutically acceptable carrier and a compound according to claim10, or a stereoisomer, a tautomer, or a pharmaceutically acceptable saltor solvate thereof.
 13. A method of treating a disease or disorder,comprising administering to a mammalian patent a compound according toclaim 1 or a pharmaceutically acceptable salt thereof, wherein saiddisease or disorder is pathological fibrosis, cancer, inflammatorydisorders, metabolic, or cholestatic disorders.
 14. The method accordingto claim 13, wherein the pathological fibrosis is liver fibrosis, renalfibrosis, biliary fibrosis, or pancreatic fibrosis.
 15. The methodaccording to claim 13, wherein said disease or disoder is nonalcoholicsteatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD),chronic kidney disease, diabetic kidney disease, primary sclerosingcholangitis (PSC), or primary biliary cirrhosis (PBC).
 16. The methodaccording to claim 13, wherein said disease or disoder is idiopathicpulmonary fibrosis (IPF).